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Immunotherapy, in particular immune checkpoint blockade (ICB) therapy targeting the programmed cell death-1 (PD-1)/programmed cell death ligand-1 (PD-L1) axis, has remarkably revolutionized cancer treatment in the clinic. Anti-PD-1/PD-L1 therapy is designed to restore the antitumor response of cytotoxic T cells (CTLs) by blocking the interaction between PD-L1 on tumour cells and PD-1 on CTLs. Nevertheless, current anti-PD-1/PD-L1 therapy suffers from poor therapeutic outcomes in a large variety of solid tumours due to insufficient tumour specificity, severe cytotoxic effects, and the occurrence of immune resistance. In recent years, nanosized drug delivery systems (NDDSs), endowed with highly efficient tumour targeting and versatility for combination therapy, have paved a new avenue for cancer immunotherapy. In this review article, we summarized the recent advances in NDDSs for anti-PD-1/PD-L1 therapy. We then discussed the challenges and further provided perspectives to promote the clinical application of NDDS-based anti-PD-1/PD-L1 therapy., (© 2022. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.)

Objective: To investigate the optimal experimental conditions (including antigen retrieval time, antibody titers and antibody incubation time) for reliable detection of programmed death-ligand 1 (PD-L1) expression using PD-L1 (22C3) antibody concentrate, and to establish a laboratory developed test for PD-L1 detection. Methods: Using Dako PD-L1 IHC 22C3 pharmDX staining procedure and scoring guidelines as the standard reference (group A), the PD-L1 expression in 25 tissue specimens (including 15 lung cancer tissues, 5 tonsil tissues and 5 placenta tissues) was detected with Flex+/HRP detection kit (EnVision) under 8 different experimental conditions (groups B1 to B8). The staining results were then compared to those in group A. Results: In group B1, 3 tissue samples showed the percentages of PD-L1 positive tumor cells were similar to those in group A, while the percentages of PD-L1 positive tumor cells were lower than those in group A in the other samples. In group B7, two case showed a positive rate higher than that in group A that was also above the positive cut-off value, and the rest of the samples had a percentage of PD-L1 positive tumor cells slightly higher than that in group A, but still below the positive cut-off value. The staining results of group B8 were the closest to those of group A compared with the other groups. Although the percentages of PD-L1 positive tumor cells in the B2 to B6 groups were decreased in various degrees as compared with group A, they were still concordant with group A's classification (positive vs. negative) and would not change the choice of clinical treatments. Conclusions: The experimental conditions are associated with detection rate of PD-L1 expression using 22C3 antibody. In the present study, the most-suitable alterative conditions in the PD-L1 detection using 22C3 antibody concentrate are those applied in the group B8 (including antigen retrieval in Dako PT Link tank at 97 ℃, pH 6.0 for 40 min and incubation with 22C3 antibodies (1∶100 dilution) at room temperature for 60 min, incubation with EnVision Flex+Linker at room temperature for 30 min, incubation with EnVision/HRP at room temperature for 30 min and DAB staining for 5 min), which could provide reliable results at minimum costs.

Objective: To study the expression of phosphates signal transducer and activator of transcription 3 (pSTAT3) and programmed death ligand-1 (PD-L1) in extranodal NK/T cell lymphomas (ENKTCL) and the relationships of pSTAT3 and PD-L1 expression with the clinicopathological characteristics and prognosis of ENKTCL. Methods: Fifty-one cases of ENKTCL diagnosed at Guangdong Provincial People's Hospital from June 2015 to February 2019 were included in the study. The expression of pSTAT3 and PD-L1 was examined using immunohistochemistry. Results: There were 35 males and 16 females, ranging from 18 to 85 years old with a median age of 47 years. The positive rates of pSTAT3 and PD-L1 expression were 68.6% (35/51) and 76.5% (39/51), respectively. pSTAT3 expression was correlated with PD-L1 expression ( P =0.033, R =0.322), while there were no associations of pSTAT3 and PD-L1 expression with the clinicopathological characteristics of ENKTCL, including age, sex, clinical site, B symptom, Ann Arbor stage, LDH value, EBV DNA load of peripheral blood and international proliferation index score. Kaplan-Meier survival analysis showed the prognoses of the pSTAT3 and PD-L1 positive groups were slightly better than the respective negative groups, but the differences were not significantly ( P >0.05). Conclusions: pSTAT3 is highly expressed in extranodal NK/T cell lymphoma and related to the expression of PD-L1, which provides a potential target and rationale for combinations of targeted therapies and immune checkpoint blockade inhibitors in the treatment of ENKTCL.

(-)-5-Epieremophilene, an epimer of the versatile sesquiterpene (+)-valencene, is an inaccessible natural product catalyzed by three sesquiterpene synthases (SmSTPSs1-3) of the Chinese medicinal herb Salvia miltiorrhiza, and its biological activity remains less explored. In this study, three metabolically engineered Escherichia coli strains were constructed for (-)-5-epieremophilene production with yields of 42.4-76.0 mg/L in shake-flask culture. Introducing an additional copy of farnesyl diphosphate synthase (FDPS) gene through fusion expression of SmSTPS1-FDPS or dividing the FDP synthetic pathway into two modules resulted in significantly improved production, and ultimately 250 mg of (-)-5-epieremophilene were achieved. Biological assay indicated that (-)-5-epieremophilene showed significant antifeedant activity against Helicoverpa armigera (EC 50 =1.25 μg/cm 2 ), a common pest of S. miltiorrhiza, implying its potential defensive role in the plant. The results provided an ideal material supply for studying other potential biological activities of (-)-5-epieremophilene, and also a strategy for manipulating terpene production in engineered E. coli using synthetic biology., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

Objectives: To investigate the clinicopathological features, therapy and prognosis of primary cardiac CD5-positive diffuse large B-cell lymphoma with C-MYC and bcl-2 double expression. Methods: Two cases diagnosed at Guangdong Provincial People's Hospital were included, the clinical data were collected; the tumor morphology, immunophenotypic profiles, therapy and prognosis were analyzed. Results: Case 1 was a 55-year-old man and case 2 was a 61-year-old women. Intraoperatively, both cases showed large masses in the right atrium or ventricle, involving adjacent tissue. Pathologically, the tumors were composed of diffusely infiltrating large lymphoid cells with high mitotic activity and apoptosis. The tumor cells were positive for CD20, CD5, bcl-6, MUM1, C-MYC and bcl-2, and the Ki-67 index was equal or greater than 90%. Case 1 had bcl-6, but not bcl-2 or MYC gene rearrangements. No MYC, bcl-2 or bcl-6 gene rearrangements were detected in case 2. Case 1 defaulted chemotherapy after operation and died 1 month after diagnosis. Case 2 was treated with 4 cycles of rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) therapy after surgery and attained partial remission, and was then treated with apatinib and ibrutinib, and remained stable 18 months after initial diagnosis. Conclusion: Primary cardiac CD5-positive diffuse large B-cell lymphoma with C-MYC and bcl-2 double expression usually shows large infiltrative mass in the right atrium or ventricle, non-germinal center like immunophenotype and high proliferation index, and this may contribute to the aggressiveness of primary cardiac lymphoma.

Objective: This study aimed to explore the potential effects of terahertz (THz) waves on primary cultured neurons from 4 rat brain regions (hippocampus, cerebral cortex, cerebellum, and brainstem) and 3 kinds of neuron-like cells (MN9D, PC12, and HT22 cells) under nonthermal conditions., Methods: THz waves with an output power of 50 (0.16 THz) and 10 (0.17 THz) mW with exposure times of 6 and 60 min were used in this study. Analysis of temperature change, neurite growth, cell membrane roughness, micromorphology, neurotransmitters and synaptic-related proteins (SYN and PSD95) was used to evaluate the potential effects., Results: Temperature increase caused by the THz wave was negligible. THz waves induced significant neurotransmitter changes in primary hippocampal, cerebellar, and brainstem neurons and in MN9D and PC12 cells. THz wave downregulated SYN expression in primary hippocampal neurons and downregulated PSD95 expression in primary cortical neurons., Conclusion: Different types of cells responded differently after THz wave exposure, and primary hippocampal and cortical neurons and MN9D cells were relatively sensitive to the THz waves. The biological effects were positively correlated with the exposure time of the THz waves., (Copyright © 2019 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.)

Objective: To analyze the prevalence, clinical characteristics and prognostic significance of the isocitrate dehydrogenase 2(IDH2) mutations in patients with acute myeloid leukemia(AML)., Methods: The bone marrow samples of 223 patients with newly diagnosed AML confirmed by MICM typing from January 2015 to October 2018 were collected. The mutation of exon 4 of IDH2 gene was detected by direct sequancing of PCR product; the incidence and types of IDH2 gene mutation in AML patients were analyzed; the clinical characteristics of AML patients with IDH2 gene mutation were analyzed and the therapeutic efficacy for these patients was evaluated., Results: In a cohort of 223 AML patients, mutations were detected in 23(10.31％) patients, among them, 15 with R140Q mutations(65.22%) , 6 with R172K mutations(26.09%) and 2 with R140W mutations(8.70%). The median age in IDH2 mutated group was older than that in non.mutated group(P=0.008). The platelet level at initial diagnosis in IDH2 mutated group was higher than that in non.mutated group(P=0.010). There was no significant statistical difference between IDH2 mutated group and non.mutated group in FAB subtypes of AML(P>0.05). But the rate of IDH2 mutation in M4 and M5 was higher. The rate of IDH2 mutations was higher in AML with normal karyotype and in AML with NPM1 mutations. R140Q mutations associated with NPM1 mutations(χ 2 =8.481，P=0.004), but R172K mutations not associated with NPM1 mutation(P>0.05). IDH2 mutated patients had a lower complete remission rate than non.mutated patients(57.14% vs 80.46%, χ 2 =5.927，P=0.015). The complete remission rate of R140Q mutated patients was not significantly statistically different from non.mutated patients. The complete remission rate of R172K mutated patients was very significantly lower than non.mutated patients(χ 2 =7.734，P=0.005). In the patients without NPM1 mutation, the 2 years overall survival in IDH2 mutated group was lower than in non.mutated group(36.36% vs 66.40%,χ 2 =3.958，P=0.047), the 2 years overall survival of R172K mutated group was significantly lower than non.mutated group(although P>0.05). In all patients, the 2 years overall survival between IDH2 mutated group and non.mutated group was not statistically different(50% vs 66.88%,P>0.05), the 2 years overall survival of R172K mutated group was significantly lower than non.mutated group(although P>0.05). In the patients with normal karyotype or with mutated NPM1, the 2 years overall survival between IDH2 mutated group and non.mutated group was not statistically different(P>0.05)., Conclusion: IDH2 gene mutations are more common in AML patients at older age, higher platelets level and normal karyotype. The rate of IDH2 mutation in M4 and M5 is higher. IDH2 gene mutations associate with NPMl gene mutations, but R172K mutations not associates with NPM1mutation. IDH2 gene mutations associate with prognosis of AML patients, R140Q mutations have no effect on prognosis of patients, but R172K mutations may be the molecular markers for poor prognosis in AML patients.

Objective: To investigate the clinicopathological features, treatment and prognosis of duodenal-type follicular lymphoma. Methods: Four cases of duodenal-type follicular lymphoma diagnosed at Guangdong General Hospital from 2014 to 2015 with detailed clinical data were included. The histomorphology, immunophenotype, treatment and prognoses were analyzed. Results: The patients' age ranged from 51 to 57 years (mean 54 years), and there were 2 males and 2 females. The involved sites were gastric fundus in one case, second portion of the duodenum in two cases and terminal ileum in one case. All patients presented with multiple mucosal granules or nodules at endoscopy. Microscopically, there were multiple mucosal neoplastic follicles, constituting grade 1-2 disease based on nodal follicular lymphoma grading system. The tumor cells were positive for CD20, CD10, bcl-6 and bcl-2. CD21 highlighted the follicular dendritic meshwork mainly at the periphery of the follicles. Proliferation index was low. Three patients received rituximab monotherapy for 4 cycles, leading to complete remission. One patient refused therapy and the disease progressed to systemic lymphoma 15 months after the initial diagnosis. Conclusions: Duodenal-type follicular lymphoma is a special variant of follicular lymphoma with indolent clinical course. The tumor exhibits morphology of low grade follicular lymphoma with characteristic dendritic meshwork at the periphery of the follicles and a low proliferation index. Prognosis is excellent. Rituximab monotherapy is treatment of choice, but a small minority of patients may progress to systemic disease.

Lead species significantly influences its mobility and bioavailability. In this study, improved BCR procedure, laboratory culture, laboratory micro X-ray fluorescence (μ-XRF), and synchrotron radiation X-ray absorption near-edge structure (XANES) were used to investigate the distribution, speciation, and bioavailability of Pb in soils from a Pb/zinc (Zn) mining area contaminated with heavy metals. The results showed that ① Pb was mainly distributed in the reducible[which binds to oxides of iron (Fe) and manganese (Mn)], acid soluble, residual, and oxidizable fractions, ② Pb content in plant tissues correlated with acid-soluble Pb content in soil, ③ μ-XRF results indicated that Pb was distributed heterogeneously, and correlated with the hot spot with high concentration of Mn. In some area, Pb also correlated with Fe, and ④ Pb was predominantly present as Pb-goethite (41%-46%), Pb 3 (PO 4 ) 2 (36%-55%), and Pb-MnO 2 (3%-24%) in soil samples. The results of this study showed that oxides of Fe and Mn, and Pb 3 (PO 4 ) 2 precipitate mainly comprise the pool of Pb in soils.

Objective: To evaluate the expression of βF1 and T cell receptor (TCR)γ in T lymphoblastic lymphoma/leukemia(T-LBL/ALL), and investigate the clinicopathological features. Methods: Fifty-one cases of T-LBL/ALL were collected at Guangdong General Hospital from 2010 to 2016, the expression of βF1 and TCRγ was assessed by immunohistochemistry. Results: There were 13 cases of children and adolescents, and 38 cases of adults. The expression rates of βF1 and TCRγ were 27.5%(14/51) and 15.7%(8/51) respectively. The proportion of adults in αβ T-LBL/ALL, TCR-silent T-LBL/ALL and γδ T-LBL/ALL was 7/14, 79.3%(23/29)and 8/8 respectively, and the difference was significant ( P =0.023). There was no statistical difference in sex, LDH, bone marrow involvement and Ann arbor stage among these three groups( P >0.05). γδ T-LBL/ALLs included 6 cases of CD4(-)/CD8(-) phenotype, whereas αβ T-LBL/ALL included 7 cases of CD4(+) /CD8(+) phenotype. There was significant difference in CD4/CD8 expression among these three groups( P <0.01). Conclusions: γδ T-LBL/ALL occurred only in adults, with predominantly CD4(-)/CD8(-) phenotype. αβ T-LBL/ALL occurred more common in children and adolescents, with predominantly CD4(+) /CD8(+) phenotype.

In the aim to realize high performance semitransparent fully coated organic solar cells, printable electrode buffer layers and top electrodes are two important key technologies. An ideal ink for the preparation of the electrode buffer layer for printed top electrodes should have good wettability and negligible solvent corrosion to the underlying layer. This work reports a novel organic-inorganic composite of phosphomolybdic acid (PMA) and PEDOT:PSS that features excellent wettability with the active layer and printed top Ag nanowires and high resistibility to solvent corrosion. This composite buffer layer can be easily deposited on a polymer surface to form a smooth, homogeneous film via spin-coating or doctor-blade coating. Through the use of this composite anode buffer layer, fully coated semitransparent devices with doctor-blade-coated functional layers and spray-coated Ag nanowire top electrodes showed the highest power conversion efficiency (PCE) of 5.01% with an excellent average visible-light transmittance (AVT) of 50.3%, demonstrating superior overall characteristics with a comparable performance to and a much higher AVT than cells based on a thermally evaporated MoO 3 /Ag/MoO 3 thin film electrode (with a PCE of 5.77% and AVT of 19.5%). The current work reports the fabrication of fully coated inverted organic solar cells by combining doctor-blade coating and spray coating and, more importantly, demonstrates that a nanocomposite of a polyoxometalate and conjugated polymer could be an excellent anode buffer layer for the fully coated polymer solar cells with favorable interfacial contact, hole extraction efficiency, and high comparability with full printing.

Bismuth nanodendrites (BiNDs) were electrodeposited on planar gallium nitride (GaN) electrode via a differential pulse voltammetric technique to fabricate the non-enzymatic hydrogen peroxide (H 2 O 2 ) sensor. SEM images revealed that the as-obtained BiNDs had numerous dendrite sub-branches, whose diameters ranged from 136 to 152nm. The BiNDs/GaN electrode showed linear amperometric responses for H 2 O 2 in the concentration range from 10µM to 1mM with the sensitivity of 60.0μAmM -1 cm -2 . Another linear range was from 1 to 10mM with the sensitivity of 23.3μAmM -1 cm -2 . The limit of detection (LOD) was 5µM with the signal-to-noise ratio of 3. The applicability of the sensor was investigated to the H 2 O 2 detection in real samples such as fetal bovine serum and milk, and the sensor exhibited excellent anti-interference capacity. The achieved results indicate that the as-prepared BiNDs/GaN sensor with good reproducibility and long-term stability was promising for detecting H 2 O 2 in practical environments., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Roads are typically man-made objects in urban areas. Road extraction from high-resolution images has important applications for urban planning and transportation development. However, due to the confusion of spectral characteristic, it is difficult to distinguish roads from other objects by merely using traditional classification methods that mainly depend on spectral information. Edge is an important feature for the identification of linear objects (e. g. , roads). The distribution patterns of edges vary greatly among different objects. It is crucial to merge edge statistical information into spectral ones. In this study, a new method that combines spectral information and edge statistical features has been proposed. First, edge detection is conducted by using self-adaptive mean-shift algorithm on the panchromatic band, which can greatly reduce pseudo-edges and noise effects. Then, edge statistical features are obtained from the edge statistical model, which measures the length and angle distribution of edges. Finally, by integrating the spectral and edge statistical features, SVM algorithm is used to classify the image and roads are ultimately extracted. A series of experiments are conducted and the results show that the overall accuracy of proposed method is 93% comparing with only 78% overall accuracy of the traditional. The results demonstrate that the proposed method is efficient and valuable for road extraction, especially on high-resolution images.

Cyanidin-3-O-glucoside (C3G), an anthocyanin belonging to the flavonoid family and commonly present in food and vegetables in human diet, has exhibited anti-inflammatory and anti-oxidant effects. This study aimed to investigate the protective ability of C3G against inflammatory and oxidative injuries, as well as to clarify the possible mechanism in lipopolysaccharide (LPS)-stimulated human umbilical vein endothelial cells (HUVECs) in vitro and acute respiratory distress syndrome mouse model in vivo. HUVECs or male Kunming mice were pretreated with C3G 1 h before LPS stimulation. C3G significantly inhibited the production of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin (IL) -6, and IL-1β) in cell supernatants and bronchoalveolar lavage fluid (BALF) as determined by enzyme-linked immunosorbent assay. Histopathologic examination with hematoxylin and eosinstaining showed that C3G pretreatment substantially suppressed inflammatory cell infiltration, alveolar wall thickening, and interstitial edemain lung tissues. C3G markedly prevented LPS-induced elevation of malondialdehyde and myeloperoxidase levels in lung tissue homogenates, wet to dry ratio of lung tissues, total cells, and inflammatory cells (neutrophils and macrophages) in BALF. Moreover, C3G reduced superoxide dismutase activity in the lung tissue homogenates. Western blot assay also showed that C3G pretreatment significantly suppressed LPS-induced activation of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways by blocking the phosphorylation of inhibitor κB-α, NF-κB/P65, extracellular signal-regulated kinase, p38, and c-Jun NH2-terminal kinase in the lung tissues. In summary, C3G may ameliorate LPS-induced injury, which results from inflammation and oxidation, by inhibiting NF-κB and MAPK pathways and playing important anti-inflammatory and anti-oxidative roles.

Cyclin D1b, a splice variant of the cell cycle regulator cyclin D1, holds oncogenic functions in human cancer. However, the mechanisms underlying cyclin D1b function remain poorly understood. Here we introduced wild-type cyclin D1a or cyclin D1b variant into non-metastatic MCF-7 cells. Our results show that ectopic expression of cyclin D1b promotes invasiveness of the cancer cells in a cyclin D1a independent manner. Specifically, cyclin D1b is found to modulate the expression of αvβ3, which characterizes the metastatic phenotype, and enhance tumor cell invasive potential in cooperating with HoxD3. Notably, cyclin D1b promotes αvβ3-mediated adhesion and invasive migration, which are associated with invasive potential of breast cancer cells. Further exploration indicates that cyclin D1b makes breast cancer cells more sensitive to toll-like receptor 4 ligand released from damaged tumor cells. These findings reveal a role of cyclin D1b as a possible mediator of αvβ3 transcription to promote tumor metastasis., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

The study is aimed to assess the genetic diversity and genetic relationship of 18 Sarcandra glabra resources from different populations,and guide parent selection of cross breeding between these resources. The molecular marker technique ISSR was used to investigate the genetic diversity of the 18 resources. Data was analyzed by POPGEN 32, and a cluster diagram was presented by UPGMA. One hundred and ninety-eight amplified fragments were obtained using 23 ISSR primers. One hundred and eighty-four polymorphic loci were identified. Nei's genetic diversity index (h) was 0.32, Shannon diversity index (I) was 0.485 4. The genetic similarity coefficient among the resources ranged from 0.383 8 to 0.878 8 in an average of 0.661 2. The genetic distance between sample S2 and sample S18 was the farthest, so as between sample S3 and sample S18 both Nei's genetic distance was 0.957 5, The genetic distance between sample S4 and sample S5 was the closest, the Nei's genetic distance was 0.129 2,and the sample S1, S2, S3, S7, S10 were significantly different from the others based on the clustering analysis, the three groups S2 vs S3, S2 vs S6, S2 vs S18 were the best parent group selection. There was a middle level of genetic differentiation in the resources. The genetic distance between resources gives useful information to guide parent selection of cross breeding.

The paper is aimed to identify SNP in Sarcandra glabra and Chloranthus spicatus, and authenticate S. glabra from Ch. spicatus and the mixture by using PCR amplification of specific alleles. SNPs in the ITS sequences of S. glabra and Ch. spicatus were found by ClustulX 2. 1 program and Bioedit software. Primers for authentic S. glabra and Ch. spicatus was designed according to the SNP site, and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system, and then optimized the PCR reaction system. Five hundred and eighty band special for S. glabra and 470 bp band special for Ch. spicatus were found by using multi-PCR reaction. The multi-PCR reaction system could be applied to identify S. glabra and Ch. spicatus's leaves.

This study was aimed to investigate the effect of SU11248 on proliferation and apoptosis of leukemia cell line K562 in vitro and its mechanism. The inhibitory effect of 3.2 µg/ml SU11248 on K562 proliferation was tested by MTT assay. The ability of SU11248 to induce apoptosis of K562 cells was examined by TUNEL and DNA ladder. The expression of C-MYC, hTERT and BCR-ABL mRNA in K562 cells was detected by RT-PCR. The protein expression of Akt and p-Akt in K562 cells was detected by Western blot. The results showed that the proliferation of K562 cells was obviously inhibited by 3.2 µg/ml SU11248 in a time-dependent manner. SU11248 could induce K562 cells apoptosis in dose-and time-dependent manner. The mRNA expression of C-MYC, hTERT and BCR-ABL was reduced significantly by SU11248 in a time-dependent manner (P < 0.05). Western blot detection showed that the expression of p-Akt protein in K562 cells decreased in dose-and time-dependent manner after SU11248 treatment, but the expression of Akt was not significantly changed. It is concluded that SU11248 can inhibit the growth of K562 cells efficiently through inducing apoptosis, its mechanism may be closely relate with the expression down-regulation of C-MYC, hTERT, BCR-ABL and the inhibition of Akt phosphorylation.

In this work, we proposed a novel simple protocol for preparing 1-aminopyrene/graphene (ApG) hybrids for fabricating label-free electrochemical impedance genosensor. Graphene, with the structure of a single-atom-thick sheet of sp(2)-bonded carbon atoms, was anchored to 1-aminopyrene (1-Ap) with the pyrenyl group viaπ-stacking interaction. The morphology, conductivity, and interaction of ApG hybrids were characterized by transmission electron microscopy (TEM), cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), UV-visible (UV-vis) and fluorescence spectra. The amino-substituted oligonucleotide probe was conjugated to 1-Ap by the cross-linker glutaraldehyde. The DNA hybridization reaction of oligonucleotide probe with target DNA was monitored by EIS. Under optimum conditions, the proposed biosensor exhibited high sensitivity and a low detection limit for detecting the complementary oligonucleotide. The target oligonucleotide could be quantified in a wide range of 1.0 × 10(-12) to 1.0 × 10(-8) M with good linearity (R = 0.9900) and low detection limit of 4.5 × 10(-13) M (S/N = 3).

NAC family is a class of transcription factors that have been typically found in plant with various functions. This type of genes plays a key regulatory role in secondary plant growth, cell division, senescence, especially in the hormone and signaling pathways. ANAC092 was reported to function in relation to lateral root development and senescence. The promoter and in situ hybridization analyses showed that ANAC092 was expressed temporally in the process of anther development. The gene was expressed in tapetum at stages 8-11 of anther development and reached the highest amount at stages 9-10, which was overlapped with the expression period of AMS (Aborted microspores). In this study, we constructed ANAC092 over-expression system, and identified homozygous transgenic lines. Compared with the wild type, the number of pollen grains in the transgenic line was decreased significantly, but the length of pollen grain was increased. qRT-PCR analysis showed that the expression of genes related to pollen development, e.g., SPL, EMS1, DYT1, and AMS was in-creased in the over-expression plants. Bioinformatics results showed that ANAC092 promoter sequence possessed seven AMS binding sites. All the results showed that ANAC092 is possibly located in the downstream of AMS and plays an im-portant role in the process of pollen development.

Qixiashan lead-zinc mine of Nanjing was one of the largest lead zinc deposits in East China Its exploitation has been over 50 years, and the environmental pollution has also been increasing. The lead concentration in the local environment was high, but lead migration and toxic mechanism has not been clear. Therefore, biogeochemistry research of the lead zinc mine was carried out. Using ICP-MS and Pb-L III edge XANES, lead concentration and speciation were analyzed respectively, and duckweed which can tolerate and enriched heavy metals was found in the pollution area. The results showed that the lead concentration of duckweed was 39.4 mg x kg(-1). XANES analysis and linear combination fit indicated that lead stearate and lead sulfide accounted for 65% and 36.9% respectively in the lead speciation of duckweed, suggesting that the main lead speciation of duckweed was sulfur-containing lead-organic acid.

The data of in vivo XRF Pb in human bone were obtained within China by using the in vivo XRF set. The K series X-ray of Pb were applied to determine the in vivo Pb concentration in tibia and calcaneus among Chinese mainland residents. For general population, the weighted average of Pb in bone was 0.4-22.7 microg x (lead g bone mineral)(-1). The uncertainty was 7.0-12.5 microg x (lead g bone mineral)(-1), and the average minimun detection limit was 20.3 microg x (lead g bone mineral)(-1). For residents from pollution area, the maximum Pb in bone reached up to 73.9 microg x (lead g bone mineral)(-1).

Objective: To investigate magnetic resonance imaging (MRI) characteristics of ovarian carcinosarcoma and the diagnostic value of MRI., Methods: The MRI features of ovarian carcinosarcoma and clinical data of 5 patients with ovarian carcinosarcoma were reviewed. All the lesions were confirmed by surgery and pathological examination., Results: MRI of ovarian carcinosarcoma in the 5 cases all showed large tumor mass and heterogeneous high-intensity on T2-weighted images and low-intensity on T1-weighted images, with laminar or stripe-like enhancement. Hemorrhage and necrosis were also displayed in some lesions. In two cases, the tumors invaded the greater omentum, sigmoid colon and the body of the uterus, with regional lymph node involvement. Pelvic effusion was observed in all the cases with pelvic hematocele in 1 case., Conclusion: MRI is useful in the detection and staging of ovarian carcinosarcoma.

Due to the impact of lead-zinc mining and smelting, the soil of lead-zinc orefield and nearby areas has been widely polluted. Moreover, the concentration of lead and zinc was high, and their concentration range was wide. Furthermore, the impact of overlapping interference between low concentration As and high Pb was serious, and the lack of soil standards of lead and zinc was also a problem. According to these problems, the present work studied an effective solution to setting up the EDXRF analysis of Pb, As, Cd, Cu and Zn of soil on lead-zinc orefield. The measurable concentration of Pb was 4.4-23 600 microg x g(-1), and that of Zn was 7.0-39 400 microg x g(-1). The detection limit was 1.1 microg x g(-1) for Pb and 0.9 microg x g(-1) for Zn. The average relative-error was 7.6% for Pb and 6.2% for Zn.

This study was purposed to explore the expression of P27(kip1)and cyclin G in patients with acute leukemia (AL) and its correlation. The reverse polymerase chain reaction (RT-PCR) was used to analyse the expression of P27(kip1) and cyclin G mRNA in 89 AL patients and 10 normal persons; Western blot was used to analyze the expression of P27(kip1) and cyclin G protein in 39 AL patients and 10 normal persons. The results showed that the cyclin G mRNA and protein expressions in new diagnosed/relapsed cases of AL were significantly higher than those in patients with remission and normal controls (p < 0.05 and p < 0.01), but there was no difference between remission cases and normal controls (p > 0.05). The expression of P27(kip1) mRNA in newly diagnosed/relapsed patients with AL was not significantly different from patients with remission and normal controls (p > 0.05), while the P27(kip1) protein expression in remission cases of AL and normal controls was significantly higher than that in new diagnosed/relapsed cases (p < 0.05), but there was no difference between remission cases and normal controls (p > 0.05). The expression of P27(kip1) negatively and lowly correlated with the expression of cyclin G in patients with AL. It is concluded that the low expression of P27(kip1) and the high expression of cyclin G in patients with AL may have some correlation with genesis and development of AL and may be an indication for poor prognosis of AL.

Objective: To study the efficacy and safety of adsorption of endotoxin and proinflammatory cytokines with HB-H-7 resin in vitro., Methods: Static adsorption experiment: HB-H-7 resin was added into plasma of endotoxin-positive patients according to the ratio of 1;10. Then, they were put in a constant temperature water bath oscillator, and oscillated for 2 hours. Before and after the experiment, plasma endotoxin, proinflammatory cytokines, protein and electrolytes were detected, and the rate of 2-hour absorption was calculated. The experiment was repeated 10 times. Dynamic adsorption experiment: 5 ml resin was put into a self-made perfusion unit. Endotoxin-positive patients plasma (50 ml) was perfused for 2 hours. The flow rate of plasma was controlled at 4 ml/min with the infusion pump, and the plasma endotoxin was determined at 0, 30, 60 and 120 minutes after plasma hemoperfusion, and the absorption rates were calculated. The results were compared with static adsorption. The influence of temperature during adsorption was determined as follows. Perfusion method was similar with dynamic adsorption experiment. The perfusion units were either placed in a 37 centigrade water path or 25 centigrade (room temperature). Then, the plasma endotoxin was measured 2 hours after plasma perfusion, and the absorption rates were calculated., Results: In static adsorption experiment, the plasma endotoxin and proinflammatory cytokines were significantly lower after adsorption with HB-H-7 resin adsorption (all P<0.05). The adsorption rates of endotoxin, tumor necrosis factor-alpha and interleukin-1, 6, 8 were 99.2%, 55.0%, 57.0%, 75.0%, 42.0%, respectively. Changes in protein were small (all P<0.05) , and there was no significant change in plasma electrolytes (all P>0.05). Dynamic adsorption rate was higher than that of static, but the differences were not significant (99.8% vs. 99.1%, P>0.05). There was no statistically significant difference between difference in temperature (37 centigrade vs. 25 centigrade, 99.8% vs. 99.9%, P>0.05)., Conclusion: HB-H-7 resin effectively absorbs endotoxin and proinflammatory cytokines in vitro. Its adsorption rate for protein is lower, and it has no obvious effects on electrolytes.

Overlapped spectra occur often in energy dispersive X-ray fluorescence spectrometry. Without a suitable curve fitting process, no correct qualitative and quantitative results can be obtained. The most difficult problems in curve fitting include that elements and their lines are unknown, background is uncertain sometimes, and initial model parameters may be in correct. In order to solve these problems, several available algorithms may be used. Among them are genetic algorithms. In the present work, polarization energy dispersive X-ray spectrometry was used in the determination of raw materials for permanent magnet. A genetic algorithm was successfully applied to the deconvolution of the overlapped spectra in magnetic materials. With its global searching capability, the genetic algorithm beatures higher resolution than the standard Marquardt-Levenberg method in resolving the overlapped X-ray spectra. Because of the powerful capability of genetic algorithms to deconvolute overlapped spectra, the algorithms are useful especially in energy dispersive X-ray spectrometry and complex material analysis.

Objective: To develop a recombinant vaccinia virus vaccine expressing HPV58 E7 and to determine its immuno-protective activity in mice bearing HPV58 E7+ tumor., Methods: E7 DNA was amplified and cloned from a plasmid containing HPV58 E7 genome by PCR. To abolish its transforming activity, the nucleotides coding for amino acid residues at positions 24 and 92 were modified by site-directed mutagenesis so that cysteine was substituted by glycine. Balb/c 3T3 cells were transfected with mE7. The expression of E7 protein by the mE7-transfected Balb/c cells was confirmed by immunofluorescence staining. The transfected cells were observed in vitro for anchorage-independent growth and tumorigenesis in nude mice. Recombinant E7 vaccinia virus vaccine was constructed by homologous recombination of HPV58 E7 vaccinia expression plasmid and vaccinia virus (Tiantan stain). The immuno-protective activity of the vaccines was determined by tumor growth inhibition and cytotoxic T lymphocytes (CTL) induction in vaccine-immunized syngeneic mice., Results: Substitution of cysteine by glycine at both positions 24 and 92 of HPV58 E7 abolished its transforming activity. Growth of HPV E7+ tumor in mice immunized with the recombinant vaccinia virus expressing HPV58 E7 was inhibited, and the surviving time of the immunized mice was prolonged. CTL activity was induced as revealed by in vitro cytotoxicity assay using E7+ tumor cells as target cells., Conclusions: HPV58 E7, with its transforming potential abolished, may be used as vaccine for immunotherapy of patients with HPV 58 related cancers.

The review focuses on the expert systems and knowledge engineering in X-ray fluorescence spectrometry. It includes mainly a knowledge-controlled strategy combining the scan-based method with the fixed channel measurements, a XRF interpretation system of spectra with fuzzy logic and pattern recognition, and an expert system for qualitative interpretation of XRF spectra using a certainty factor. In the review, a series of the studies of exploring the knowledge engineering system in XRF are also included, which consists of four parts, i.e. spectra identification, pattern recognition with decision-making, quantitative determination combined with the theoretical alpha coefficients and neural networks, and XRF analysis without standards.

gamma-Interferon (IFN-gamma) was detected by ELISA assay in ascitic fluid from a number of ovarian cancer patients. To study its clinical significance, the effect of IFN-gamma on the metastatic potential of a mouse mammary adenocarcinoma, MA-891, was explored. Pretreatment of the tumor cells in vitro for 48hr with recombinant INF-gamma significantly increased the number of lung tumor nodules after i. v. or s. c. inoculation into (TA2 x 615) F1 mice. In contrast, when recombinant IFN-alpha pretreated MA-891 cells were likewise injected into mice significant decrease in metastatic potential was seen. The study in vitro indicated that pretreatment of the tumor cells with IFN-gamma but not with IFN alpha resulted in a decrease in susceptibility to NK cell cytotoxicity. In as much as both IFN-alpha and IFN-gamma can induce MHC class I expression on target cells. The increase in metastatic potential of IFN-gamma-treated tumor cells can be explained only partially on the basis of their reduced NK cells susceptibility.

Natural killer cells/large granular lymphocytes (NK/LGL) separated on discontinuous Percoll gradient from rat spleen cells were injected iv to rats (7 x 10(7) cells/rat) 3 days following iv inoculation of 2 x 10(6) Walker-256 cells. Two and 4 hr after NK/LCL injection, animals were sacrificed and the lungs examined by light and immunoelectron microscopy. The latter was done using colloidal gold-labelled polyclonal antibody against purified rat LGL cytoplasmic granules. At 2 hr following iv NK/LGL, in addition to the scattered individual tumor cells and minute tumor foci, many lymphocytes were seen accumulating in the small pulmonary vessels and capillaries. This was not observed in tumor-inoculated control rats. At 4 hr, many extravasated lymphocytes reached the lung parenchyma, some of which had attached to the tumor cells. Immunoelectromicroscopically, lymphocytes were found in intimate contact with the tumor cells with the cytoplasmic gold particles clustering at the cell contact site. Gold particles could also be seen closely adherent to the plasma membrane of degenerating tumor cells. This is the first in vivo demonstration of the role of cytotoxic granules of NK cells in tumor cell lysis.

NAD glycohydrolase (NADase) (E.C. 3.2.2.5) from five-pace snake (Agkistrodon acutus) venom was purified to electrophoretic homogeneity through a 4-step isolation procedure, including column chromatography using DEAE-Sephadex A-50, Sephadex G-75, CM Sephadex C-50 and Sephadex G-100. The final product was 11.8-fold purified with a 3.9% yield. The pure enzyme showed maximal activity at about 40 degrees C with optimal pH at 7.5. It was a glycoprotein with a pI of 7.6. Its mol. wt was respectively 98,000 as measured by gel filtration and 50,000, by SDS-PAGE. There was only one N-terminal residue, proline. NADase is thus composed of two identical subunits in each molecule. The enzyme contained copper ions. NADase activity was lost when the copper enzyme complex was treated with EDTA. The Km of the enzyme for beta-NAD, NADP and beta-NGP were 0.50 mM, 0.13 mM and 0.16 mM respectively.

We have isolated genomic and cDNA clones for a Drosophila gene resembling the human beta-amyloid precursor protein (APP). This gene produces a nervous system-enriched 6.5-kilobase transcript. Sequencing of cDNAs derived from the 6.5-kilobase transcript predicts an 886-amino acid polypeptide. This polypeptide contains a putative transmembrane domain and exhibits strong sequence similarity to cytoplasmic and extracellular regions of the human beta-amyloid precursor protein. There is a high probability that this Drosophila gene corresponds to the essential Drosophila locus vnd, a gene required for embryonic nervous system development.