Anoikis-related gene signature is associated with immune infiltration and predicts the prognosis of non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is the most common histological type of lung cancer. With the in-depth exploration of cell death manners, numerous studies found that anoikis is an important mechanism that associated with treatment. Therefore, we aimed to explore the prognostic value and treatment guidance of anoikis in NSCLC patients. In the current study, we first constructed a prognostic model based on the anoikis-related genes based on bulk RNA-sequencing and single-cell RNA-sequencing (scRNA-seq) dataset. Then, immuno-correlations of anoikis-related risk scores (ARGRS) were analyzed. In addition, HMGA1, a risky gene in ARGRS, was further explored to define its expression and immuno-correlation. Results showed that patients with higher ARGRS had worse clinical outcomes. Moreover, the five genes in the prognostic model were all highly expressed on tumor cells. Moreover, further analysis found that the ARGRS was negatively correlated with ImmuneScore, but positively with tumor purity. Besides, patients in the ARGRS-high group had lower levels of immunological characteristics, such as the immune-related signaling pathways and subpopulations. Additionally, in the immunotherapy cohorts, patients with the ARGRS-high phenotype were more resistant to immunotherapy and tended to not achieve remission after treatment. Last, HMGA1 was chosen as the representative biomarker, and analysis of the in-house cohort showed that HMGA1 was highly expressed in tumor tissues and correlated with decreased T cell infiltration. To sum up, ARGRS was correlated with a desert tumor microenvironment and identified immune-cold tumors, which can be a novel biomarker for the recognition of immunological characteristics and an immunotherapeutic response in NSCLC.

Towards fast sensing along ultralong BOTDA: flatness enhancement by utilizing injection-locked dual-bandwidth probe wave.

Brillouin optical time-domain analysis (BOTDA) using distributed Brillouin amplification (DBA) only requires a milliwatt-level pump to achieve a sensing range beyond 100 km, which provides a powerful tool for temperature/strain sensing. However, similar to the majority of other long-range BOTDAs, the state-of-the-art reports require > 1000 times average, severely restricting the sensing speed. The blind area over tens of kilometers caused by the nonuniform Brillouin response and parasitic amplitude modulation (AM) are crucial factors affecting the signal-to-noise ratio (SNR). Here, a comprehensive performance optimization and substantial enhancement for BOTDA sensors was presented by the direct demodulation of an injection-locked dual-bandwidth probe wave. Injection locking (IL) can completely eliminate the impact of AM noise; dual-bandwidth probe enables self-adaptive pulse loss compensation, thereby intensifying the SNR flatness along the ultralong fiber, and direct probe demodulation can overcome nonlocal effects and allows ∼19.7 dB enhancement of probe input power. Therefore, using only 100 times average, ∼148.3 km sensing, and ∼5 m spatial resolution were achieved with < ∼0.8 MHz standard deviation of Brillouin frequency shift (BFS) over a broad range (∼131.7 km). The reduction in averages was more than 10 times that of the reported majority of long-range BOTDAs. Such performances were achieved without using time-consuming or post-processing techniques, such as optical pulse coding and image denoising. Because this approach is compatible with optical chirp chain technique without frequency sweeping, fast acquisition (0.3 s) was also realized, which has the potential for fast sensing at 3.3 Hz along a ∼150 km fiber.

MicroRNAs and Transcripts Associated with an Early Ripening Mutant of Pomelo (Citrus grandis Osbeck).

'Liuyuezaoyou' is an early-ripening cultivar selected from a bud mutation of Citrus grandis Osbeck 'Guanximiyou'. They were designated here as MT and WT, respectively. The fruit of MT matures about 45 days earlier than WT, which was accompanied by significant changes in key phytohormones, sugar compounds and organic acids. Recent studies have showed that microRNAs (miRNAs) play an important role in regulation of fruit ripening process. The aim of this study was to compare MT fruits with WT ones to uncover if miRNAs were implicated in the ripening of C. grandis. Fruits of both WT and MT at four developmental stages were analyzed using high-throughput sequencing and RT-PCR. Several independent miRNA libraries were constructed and sequenced. A total of 747 known miRNAs were identified and 99 novel miRNAs were predicted across all libraries. The novel miRNAs were found to have hairpin structures and possess star sequences. These results showed that transcriptome and miRNAs are substantially involved in a complex and comprehensive network in regulation of fruit ripening of this species. Further analysis of the network model revealed intricate interactions of miRNAs with mRNAs during the fleshy fruit ripening process. Several identified miRNAs have potential targets. These include auxin-responsive protein IAA9, sucrose synthase 3, V-type proton ATPase, NCED1 (ABA biosynthesis) and PL1/5 (pectate lyase genes), as well as NAC100 putative coordinated regulation networks, whose interactions with respective miRNAs may contribute significantly to fruit ripening of C. grandis.

The complete chloroplast genome sequence of Citrus maxima (Burm.) Merr. 'Guanximiyou'.

Citrus maxima (Burm.) Merr. 'Guanximiyou' is a major citrus tree largely cultivated in China. A previous study has reported the complete chloroplast genome of C. maxima, but there may be some differences between wild species and cultivating variety. In this study, the complete chloroplast genome sequence of 'Guanximiyou' pummelo was characterized using BGISEQ-500 sequencing. The chloroplast genome was 160,186 bp in length and separated into four distinct regions such as large single-copy region (87,939 bp), small single-copy region (18,395 bp), and a pair of inverted repeat regions (26,926 bp). The genome contained a total of 109 genes including 79 protein-coding genes, 29 tRNA genes, and 4 rRNA genes. Phylogenetic maximum-likelihood analysis revealed that 'Guanximiyou' pummelo was clustered with other Rutaceae species with high bootstrap values.

The role of miR-7 as a potential switch in the mouse hypothalamus-pituitary-ovary axis through regulation of gonadotropins.

The hypothalamus-pituitary-ovary (HPO) axis plays fundamental roles in female neuroendocrinology and reproduction. Pituitary gonadotropins are located in the center of this axis. Previous investigation suggested that miR-7 is closely linked with gonadotropins. However, the interaction between miR-7 and the HPO axis remains unclear. This study aims to determine whether and how miR-7 functions in this axis. A mouse ovariectomy model and mouse primary pituitary cells were used in this study. The results showed that miR-7 is localized to gonadotrophs and somatotrophs. miR-7 can inhibit the expression, synthesis and secretion of gonadotropins, but not growth hormones. Gonadotropin-releasing hormone (GnRH) has inhibitory effects on miR-7, while estrogen enhances miR-7 expression. miR-7 is vital for the pathway by which GnRH and estrogen regulate gonadotropins by targeting v-raf-leukemia viral oncogene 1 (Raf1). Together, these results indicate that miR-7 acts as a potential switch in the feedback loop of the HPO axis by regulating gonadotropins.

The influence mechanism of bioclogging on pollution removal efficiency of vertical flow constructed wetland.

The effect of change of hydraulic characteristic and microbial community on pollution removal efficiency of the infiltration systems in the bioclogging development process remain poorly understood. In this study, therefore, the pollutant removal as a response to hydraulic conductivity reduction and the change of diversity and structure of microbial communities in vertical flow constructed wetlands (VFCWs) was investigated. The results indicated that the richness and diversity of the bacterial communities in the columns at different depths were decreased, and the microbial communities of the genus level were changed in the process of bioclogging. However, the variation of microbial communities has a low impact on the purification performance of VFCWs because the abundance of function groups, respiratory activity, and degradation potentiality of microorganisms remain steady or even get improved in the columns after bioclogging. On the contrary, the hydraulic efficiency of VFCWs decreased greatly by 16.9%, 9.9%, and 57.1% for VFCWs filled with zeolite (Column I), gravel (Column II), and ceramsite (Column III), respectively. The existence of short-circuiting and dead zones in the filter media cause the poor pollution removal efficiency of VFCWs due to the short contact time and decrease of oxygenation renewal, as well as low activity in the dead zone.

Behavior of selected organophosphate flame retardants (OPFRs) and their influence on rhizospheric microorganisms after short-term exposure in integrated vertical-flow constructed wetlands (IVCWs).

Considering its ubiquitous occurrence and potential adverse effects of organophosphorus flame retardant (OPFR), it is urgent to explore the efficient treatment for OPFRs wastewater. Thus, integrated vertical-flow constructed wetlands (IVCWs) were set up to comparatively evaluate their nitrogen removal capacity under tidal flow operations and to investigate environmental behavior and rhizosphere microbial responses after short-term exposure to three OPFRs. The results show that IVCWs have an excellent TN removal rate (628.13 ± 110.63 mg m-2 d-1) and moderate mitigation efficiencies (48.37 ± 9.52 to 82.28 ± 7.48%) for target OPFRs when treating low-C/N ratio wastewater. Moreover, the sorption of selected OPFRs to soil (28.85-308.41 ng g-1, dry weight (dw)), igneous rock (659.85-970.80 ng g-1 dw) and zeolite (1045.60-1351.70 ng g-1 dw) and accumulation in tissues of C. alternifolius (0-289.68 ng g-1 dw) and P. australis (0.56-108.22 ng g-1 dw) showed a hydrophobicity-specific feature. Namely, the highly hydrophobic compound tricresyl phosphate (TCrP) partitioned preferentially to sediment, and the chlorinated analytes were more easily taken up and then translocated into the plant body. Simultaneously, further mass balance analysis revealed the fate of OPFRs in IVCW components. A total of 53.25% of the highly hydrophobic TCrP inflow mass settled in sediment, while tris (2-chloroethyl) phosphate (TCEP) and tris (1-chloro-2-propyl) phosphate (TCPP) were more liable to discharge (35.33-50.89%) and other pathways (38.77-39.87%). Furthermore, the abundance of aerobic denitrifying bacteria (AD) in rhizosphere soil (2.25-5.12%), jointly with the prevalence of nitrobacteria (NOBs, 1.84-13.60%) and denitrifying bacteria (DNBs, 5.84-7.89%) in sublayer matrices, was responsible for superior TN removal. Additionally, the rhizosphere microbial richness, diversity and nitrogen-related microorganisms were clearly influenced by the presence of OPFRs. Notably, the genera Pseudomonas and Sphingobium might be the functional microorganisms for mixture OPFRs biodegradation.

Identification of Appropriate Reference Genes for Normalizing miRNA Expression in Citrus Infected by Xanthomonas citri subsp. citri.

MicroRNAs (miRNAs) are short noncoding RNA molecules that regulate gene expression at the posttranscriptional level. Reverse transcription-quantitative PCR (RT-qPCR) is one of the most common methods used for quantification of miRNA expression, and the levels of expression are normalized by comparing with reference genes. Thus, the selection of reference genes is critically important for accurate quantification. The present study was intended to identify appropriate miRNA reference genes for normalizing the level of miRNA expression in Citrus sinensis L. Osbeck and Citrus reticulata Blanco infected by Xanthomonas citri subsp. citri, which caused citrus canker disease. Five algorithms (Delta Ct, geNorm, NormFinder, BestKeeper and RefFinder) were used for screening reference genes, and two quantification approaches, poly(A) extension RT-qPCR and stem-loop RT-qPCR, were used to determine the most appropriate method for detecting expression patterns of miRNA. An overall comprehensive ranking output derived from the multi-algorithms showed that poly(A)-tailed miR162-3p/miR472 were the best reference gene combination for miRNA RT-qPCR normalization in citrus canker research. Candidate reference gene expression profiles determined by poly(A) RT-qPCR were more consistent in the two citrus species. To the best of our knowledge, this is the first systematic comparison of two miRNA quantification methods for evaluating reference genes. These results highlight the importance of rigorously assessing candidate reference genes and clarify some contradictory results in miRNA research on citrus.

Characterization of the complete chloroplast genome of the Hongkong kumquat (Fortunella hindsii Swingle).

Hongkong qumquat (Fortunella hindsii Swingle) is a wild citrus species native to China. In this study, we firstly reporteded its complete chloroplast genome using BGISEQ-500 sequencing. The chloroplast genome is 160,145 bp in size, containing a large single copy region (87,467 bp), a small single copy region (18,730 bp), and a pair of IR regions (26,974 bp). The chloroplast genome contains 112 unique genes, including 79 protein-coding genes, 29 tRNAs, and 4 rRNAs. Phylogenetic maximum-likelihood analysis indicated that F. hindsii is closely related to Citrus species. The complete chloroplast genome would be subsequently used for citrus species researches.

A review on removal of organophosphorus pesticides in constructed wetland: Performance, mechanism and influencing factors.

The residues of organophosphorus pesticides (OPPs) have been widely detected in rivers, the gulf, and even groundwater and drinking water, which may pose a serious threat to aquatic ecosystems and human health. Compared to other treatments, constructed wetlands (CWs) have been demonstrated to be a cost-effective alternative risk mitigation strategy for non-point-source pesticide pollution. This review summarizes 32 studies related to the remediation of OPPs in 117 CWs during 2001-2017 worldwide. The performances, mechanisms and influencing factors in the studies are comprehensively and critically reviewed in this paper. Overall, the OPPs were efficiently removed with an efficiency up to 87.22 ±â€¯16.61%. The removal efficiency, differences and related reasons among different types of CWs in developed and developing countries and the different types of OPPs in CWs are well-evaluated in detail. In addition, the main processes for OPPs removal in CWs involve phytoremediation (plant uptake, phytoaccumulation, phytovolatilization and phytodegradation), substrate adsorption or sedimentation, and biodegradation. Based on the quantitative analysis by mass balance, for water-soluble pesticides, the dominant removal process was via microbiological degradation. This result was in contrast to findings obtained with hydrophobic OPPs, for which the dominant processes were biodegradation and sorption by substrate. Therefore, the behavior of microbial transformation prevails. Additionally, the presence of plants can facilitate the elimination of OPPs in CWs, promoting the process by an average percentage of approximately 6.19 ±â€¯9.46%. Statistical analysis shows that loading of inlet OPPs is the largest limiting factor and that the HRT and T are the most significant parameters that influence the efficiency of trapping OPPs in CWs. Simultaneously, we can also obtain suitable parameters for the design and operation of CWs. This review promotes further research on plant-microbe joint combined remediation and examines the different behaviors of water-soluble and hydrophobic OPPs in CWs.

[Effects of CANELIM capsule drug-contained serum on viscoelasticity and adhesive mechanical properties of cholangiocarcinoma cells].

By way of gastrogavage, we administered CANELIM capsules to rat for prepering the drug-contained serums. And then the serums obtained were used to plant cholangiocarcinoma cells. Lastly, using the micropipette aspiration technique, we investigated the effects which the drug-contained serums of different doses have on the viscoelasticity and adhesive mechanical properties of cholangiocarcinoma cells. The results showed that cholangiocarcinoma cells presented a characteristic of high elastic coefficient and low viscous coefficient. After being treated by the high dose and middle dose drug-contained serums, the viscoelastical properties of cholangiocarcinoma cells K1, K2 and micro evidently decreased (P < 0.01). But the properties of low dose did not evidently change. The adhesive force between cholangiocarcinoma cells and CD44v6 protein significantly reduced with the increasing of the dose of CANELIM capsules (P < 0.01). It is suggested that CANELIM capsules would destroy the cytoskeleton of cholangiocarcinoma cells, restrain the adhesion molecule CD44v6 on membrane from expressing, reduce the adhesion probability between cholangiocarcinoma cells and vasal endothelial cells, and finally, prevent the metastasis of cholangiocarcinoma cells.

[Expression of cyclin A1 mRNA in patients with myelodysplastic syndrome and its clinical significance].

The purpose of this study was to evaluate the expression of cyclin A1 mRNA in patients with myelodysplastic syndrome (MDS) and its clinical significance. The expression of cyclin A1, cdk2 and p21(cip1) mRNA in the bone marrow from 56 patients with MDS and 10 normal control were measured by using reverse transcription polymerase chain reaction (RT-PCR) technique. The results indicated that the positive rate and the expression level of cyclin A1 in MDS patients (69.64%; 0.964 +/- 1.879) were significantly higher than those in normal control (0%; 0.012 +/- 0.014) (p < 0.01). Among de-novo MDS patients, the expression level of cyclin A1 mRNA in the MDS-RAEB group (1.895 +/- 1.769) was higher than that in MDS-RA group (0.629 +/- 1.583) (p < 0.01). The expression level of cyclin A1 mRNA in post-treatment group was significantly lower than that in prior-treatment group (p < 0.01). It is concluded that the mRNA expression of cyclin A1 in MDS patients is higher than that in normal control, the abnormal expression of cyclin A1 may be used as a prognostic marker in MDS patients.

[Establishment of a mechanical experimental system of biologic tract tissue with auto-measurement based on measuring mark].

As the core of mechanical experimental system of biologic tract tissue, the automatic measuring program of serial images based on measuring mark could provide two basic parameters for mechanical analysis, namely tissue length and average external diameter. Hybrid programming between VC+ + and Matlab is conducted. Subprogram will be in charge of the processing of single image (color-gray transform, image segmentation, pick-up target area according to measuring mark), while the main program written by VC+ + will orderly call the above subprogram again and again when it is traversing through the image sequence which records the process of a tissue's expansion and constriction under force, so the automatic measure of the serial images and mechanical analysis is achieved. The results showed: the experimental system could avoid the contrived errors caused by naked eye identification and manual choosing measuring area; the measuring precision could satisfy the need of tract tissue mechanical analysis; the system could save time and energy dramatically. The fact of tract tissue accords with the applicable conditions of mechanical analysis theory used in our experimentations.

[The clinical value of the 99Tcm-MIBI imaging in predicting the prognosis of malignant lymphoma].

OBJECTIVE: To predict the chemotherapy response in Malignant lymphoma (ML) using 99Tcm-MIBI imaging, and to evaluate whether 99Tcm-MIBI scintigraphy parameters may have a precise predictive value in the expression of MDR1 and multidrug resistance-related-protein genes. METHODS: Twenty-three patients with histologically proved Malignant lymphoma underwent 99Tcm-MIBI scintigraphy before chemotherapy. Tumor-to-background [corrected] (T/B) ratios of both early (10 min) and late images (1 h) and the percentage rate of washout (WR%) were measured; The mRNA expressions of MDR1 and MRP were measured by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR); therapeutic reaction was evaluated by clinical and radiologic methods after completing 6 - 8 cycle chemotherapy with/without involved field radiotherapy for large tumors. RESULTS: The WR% (16 +/- 6) of both MDR1 and MRP simultaneously negative expression group was significance lower than both simultaneously positive expression group (33 +/- 5), and also lower than either MDR1 or MRP positive expression group (28 +/- 6) (both P < 0.01); There was no significant difference between the both simultaneously positive expression group and either MDR1 or MRP positive expression group (P = 0.26). The early images, late images T/B ratios and WR% of MDR1 positive group were 3.0 +/- 1.1, 2.5 +/- 0.8 and 17 +/- 7 respectively; MDR1 negative group were 3.4 +/- 1.0, 2.3 +/- 0.7 and 32 +/- 6 respectively. There were no significant difference between the MDR1 positive group and MDR1 negative group in either early images or late images T/B ratios (P > 0.05), but the WR% was significant different between them (P < 0.01). The early images, late images T/B ratios and WR% of MRP positive group were 3.1 +/- 1.2, 2.5 +/- 0.8 and 19 +/- 8 respectively; MRP negative group were 3.3 +/- 1.0, 2.3 +/- 0.7 and 31 +/- 6 respectively. The WR% of MRP positive group was significantly higher than that of MRP negative group (P = 0.003); T/B ratios of both early and late images were all significantly different between them (P = 0.72, P = 0.60). Both levels of MDR1 and WR% were significantly positively correlated with therapeutic response (both P < 0.05), but there was no significant correlation between levels of MRP and therapeutic response (P = 0.052). CONCLUSION: As a untraumatic imagology instrument, 99Tcm-MIBI can accurately reflect the expression and functional status of MDR1 and MRP, and can predict the therapeutic response of ML, thus could be used for individualized treatment planning. 99Tcm-MIBI would be benefit to ML patients.

[Expression of midkine in patients with acute myeloid leukemia and its significance].

The study was aimed to investigate the expression of midkine (MK) in bone marrow mononuclear cells (BM MNC) from 65 acute myeloid leukemia patients and 15 normal controls. The method of RT-PCR was used to examine the expression of MK mRNA in BM MNC. Parts of samples were incubated for 24 hours and the gene expression of MK in the BM MNC was detected by means of Western blot. The results showed that the expression of MK of BM MNCs in 50 newly diagnosed AML patients (0.331 +/- 0.436) and 15 AML patients in relapse (0.374 +/- 0.463) were markedly higher than that in 15 CR cases (0.067 +/- 0.190), and 15 normal controls (0), respectively. The complete remission in MK positive patients (63.16%) was significantly lower than that in MK negative group (93.55%). The patients with positive MK expression had a higher relapse rate than those with negative MK expression. The positive rate of MK gene expression in drug-resistant patients and drug-sensitive patients were 57.69% and 25.64% respectively and there was positive correlation between the gene expressions of MK and bcl-2 (P < 0.01) (r = 0.0556, P < 0.001). It is concluded that MK can be secreted by AML cells and involved in drug-resistant, its positive expression may be associated with the poor prognosis in newly diagnosed AML patients. The inhibitory effect of MK on apoptosis of leukemic cells is induced by upregulating bcl-2 expression.

Expressions of cyclin E2 and survivin in acute leukemia and their correlation.

Cyclin E2 is present in solid tumors, while its expression and clinical value in acute leukemia is unknown. This study was aimed to investigate the expression of cyclin E2 and survivin gene in bone marrow cells from patients with acute leukemia and their relationship. Reverse transcription polymerase chain reaction was used for detection of the expression of cyclin E2 and survivin mRNA in 84 adult patients with acute leukemia which included 16 cases of relapse, 60 cases of de novo acute leukemia, 8 cases of continuously complete remission, and 20 normal persons as controls. The results showed that (1) positive expression of cyclin E2 (70.24%) in acute leukemia patients was significantly higher than that (0%) in controls, positive expression of survivin (72.62%) in acute leukemia patients was higher than that (30%) in control. (2) the expression of cyclin E2 positively correlated with that of survivin in acute leukemia patients. (3) remission rate in cyclin E2-positive patients (55.81%) was lower than that (88.24%) in cyclin E2-negative patients, the rate of cyclin E2 expression in relapse group was the highest among the three groups; while that in continuously complete remission group was the lowest among the three groups. (4) positive rate of cyclin E2 expression (59.32%) in patients with acute myelocytic leukemia was lower than that (96%) in patients with acute lymphocytic leukemia, no correlation between cyclin E2 expression and white blood cell counts of patients was found. It is concluded that the overexpression of cyclin E2 has been confirmed for the first time to positively correlate with the expression of the survivin in acute leukemia patients, and implicate the poor prognosis. Cyclin E2 may be used as a marker for examination of minimal residual disease.

[Relationship between VEGF and MMP-2, MMP-9 in 82 patients with acute myeloid leukemia].

In order to investigate the relationship between VEGF and matrix metalloproteinase (MMP)-2, -9 in acute myeloid leukemia patients, and evaluate the significance of them in extramedullary leukemic invasion, the expressions of MMP-2 mRNA, MMP-9 mRNA, VEGF mRNA in bone marrow from 86 patients with acute myeloid leukemia (AML), as well as human hematopoietic cell lines were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The proteolytic activities of MMP-2 and MMP-9 in the supernatants were measured by zymography. The VEGF protein in serum of all samples was detected by ELISA. All these results were analyzed for determination of the relationship between VEGF and MMP-2, MMP-9. The results showed that there was a positive correlation between expressions of MMP-2 mRNA or MMP-9 mRNA and VEGF mRNA or protein. But no such correlation was demonstrated in the AML (CR) and normal control (NC) groups. A higher expression level of MMP-2 and MMP-9 in the VEGF positive group was found, as compared with the negative group (P < 0.05). More extramedullary infiltration occurred in VEGF positive groups than that in VEGF negative groups of AML. The expression of bcl-2 in HL-60 cells was upregulated by VEGF. It is concluded that there are significantly positive correlations between the expression of MMP-2 and MMP-9 with VEGF mRNA or protein levels in AML patients. VEGF can upregulate the expression of MMP-2, MMP-9 in HL-60 and a part of the primary leukemic cells. VEGF and MMP-2, MMP-9 may participate in the extramedullary leukemic invasion of AML patients.

Expressions of cyclin B1 and p21cipl in adult acute leukemia and their correlation.

Cyclin B1, a positive regulator, controls mitosis occurrence, plays an important role in cell proliferation. To investigate the clinical significance of cyclin B1, the expression of cyclin B1 in acute leukemia (AL) patients was measured; the expression of cyclin B1 and p21(cipl), and their cell cycle distribution were assayed by flow cytometry in 136 adult patients with newly diagnosed AL, 10 continuous complete remission (CCR) AL and 17 normal controls; the mRNA of cyclin B1 and p21(cipl), and the proliferation cell nuclear antigen (PCNA) in patients and normal controls were detected with semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). The results showed that the expression of cyclin B1 in newly diagnosed AL patients was significantly higher than that in normal controls. For the relapsed AL patients, the cyclin B1 expression was also higher than that in normal controls, but lower than that in newly diagnosed cases, there was no significant difference between the remission cases and normal controls, nor difference between CCR AL patients and normal controls. All patients with high cyclin B1 expression had an unscheduled expression manner, that cyclin B1 protein appeared in G(1) phase, and in some case it even higher than that of G(2) phase. The response rate (partial remission + complete remission) and survival rate in the cyclin B1 high expressed patients were higher than that of cyclin B1 low expressed patients. The relapse rate in cyclin B1 high expressed patients was higher than that in cyclin B1 normally expressed patients. The survival rate in cyclin B1 high expressed patients was higher than that in cyclin B1 low expression patients. A negative correlation between the expression of cyclin B1 and p21(cipl) was observed. Additionally, cyclin B1 protein expression was generally correlated with proliferation index (PI) and proliferation cell nuclear antigen (PCNA). It is concluded that this study demonstrates for the first time cyclin B1 overexpression and abnormally distribution in cell cyclin of newly diagnosed AL patients. It was considered that cyclin B1 may play an important role in leukemic pathogeneses and can be one of the factors influencing the prognosis of AL patients.

[Synergetic cytotoxicity of cisplatin and etoposide to leukemia cell line K562 and its mechanism].

BACKGROUND & OBJECTIVE: Etoposide (VP-16) is one of the most common chemotherapy drugs, but its usage is limited by drug resistance. Some researches on solid tumors show that cisplatin (DDP) have synergetic effect with VP-16. This study was to evaluate synergetic cytotoxicity of VP-16 and DDP to leukemia cell line K562, and explore the mechanism. METHODS: K562 cells were treated with VP-16 (0 or 5 microg/ml) and DDP (0, 0.3, 3, 15, or 30 microg/ml) in different combination patterns. Inhibitory effect of VP-16 and DDP on survival of K562 cells was measured by MTT assay. Cell apoptosis was evaluated by AO/EB double fluorescent labeling. The expression of topoisomerase (TOPO) II alpha and II beta, and transcription factor Sp1 and Sp3 were measured by semi-quantitive reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. RESULTS: MTT assay showed significant synergetic cytotoxicity of VP-16 and DDP. VP-16 in combination with DDP obviously enhanced cell apoptosis. RT-PCR showed that DDP significantly up-regulated the expression of TOPO II and Sp1 in concentration-dependent manners (TOPO II alpha, II beta, and Sp1 were up-regulated by 36%, 25%, and 75% of control, respectively, when treated with 30 microg/ml of DDP), and down-regulated Sp3 by 49% of control; VP-16 (5 microg/ml) down-regulated TOPO II alpha by 71.9%, and up-regulated Sp3 by 14%; VP-16 (5 microg/ml) in combination with DDP (30 microg/ml) up-regulated TOPO II alpha by 83%, II beta by 11%, and Sp1 by 58% when compared with using VP-16 alone (but the levels were lower than using DDP alone), and down-regulated Sp3 by 61.3% when compared with using DDP alone. Western blot showed similar results to RT-PCR. CONCLUSION: Through up-regulating TOPO II, DDP could enhance the chemotherapeutic effect of VP-16 on K562 cells by provide more target enzyme to act on.

[Expression of cyclin g2 mRNA in patients with acute leukemia and its clinical significance].

To evaluate the expression of cyclin G2 mRNA in patients with acute leukaemia (AL) and its clinical value, the expression of cyclin G2, G1 and P53 mRNA in the bone marrow from 74 AL patients and 10 normal individuals as control were detected with reverse transcription polymerase chain reaction (RT-PCR). The positive segment of cyclin G2 was analyzed by DNA sequencing. The results showed that (1) the positive rate and the expressing level of cyclin G2 in AL patients (52.7%, 0.552 +/- 0.498) were significantly lower than those in normal control (100%, 1.953 +/- 0.675) (P < 0.01); (2) among new diagnosed AL patients, the complete remission (CR) rate (69.2%) in the positive cyclin G2 patients was higher than that (40%) in negative cyclin G2 patients (P < 0.05); (3) the positive rate of cyclin G2 (43.6%) in resistance group was significantly higher than that (68.6%) in sensitive group (P < 0.01); (4) following-up for 14.3 month (11 - 18.5 month) in 28 AL patients with CR, there were 10 relapsed in 11 AL patients with low expression level of cyclin G2 (90.9%); and 7 relapsed in 17 AL patients with high expression (41.2%), and there was significant difference (P < 0.05). In conclusion, the expression of cyclin G2 in AL patients was higher than that in normal control, the abnormal expression of cyclin G2 might be a prognostic marker of CR in AL patients.