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 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.

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.

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.

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.

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.

[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.

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.

[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.

Correlation between cyclin A gene expression in adult patients with acute leukemia and drug resistance.

In order to investigate the relationship between the expression of cyclin A and drug resistance in adult patients with acute leukemia (AL), the mRNA expression of cyclin A, mdr1, Top II alpha, bcl-2 was detected in 64 adult patients with AL and 20 normal controls by semi-reverse transcription polymerase chain reaction (semi-RT-PCR). It was found that the cyclin A and Top II alpha mRNA expression levels in drug resistant group were significantly lower than in sensitive group (P < 0.01). Under the same experimental condition no cyclin A mRNA expression was detectable in all normal controls. The mdr1 and bcl-2 mRNA expression levels in resistant group were significantly higher than in sensitive group (P < 0.01), cyclin A and Top II alpha gene expression levels were closely correlated (rs = +0.794, P = 0.000, n = 64) in all AL patients, but cyclin A was not correlated with mdr1 and bcl-2 gene expression levels. In drug resistant group there was a negative correlation between the gene expression levels of cyclin A and mdr1 (rs = -0.337, P = 0.029). The 10 AL patients with positive lower expression of both cyclin A and Top II alpha were all resistant to drugs. Logistic regression of Binary analysis showed the correlation between the lower expression of cyclin A and drug resistance. It was concluded that lower expression of cyclin A gene might be an unfavorable prognostic factor for patients with AL, and detection of both cyclin A and Top II alpha gene expression would predict drug resistance in AL patients.

[Effect of arsenic trioxide on the expression of cyclins gene in HL60 cells].

OBJECTIVE: To investigate the expression of cyclins related gene in HL60 cells before and after arsenic trioxide treatment by gene chip. METHODS: Total mRNAs were extracted from untreated or arsenic trioxide treated HL60 cells. Then two cDNA probes were made from these two mRNAs which were labeled by fluoro link Cy3-ducpp deoxyuridine triphosphate (Cy3-dUTP) or Cy5-dUTP fluorescence dyes respectively, hybridized with gene chip and scanned for fluorescent intensity. Different expression genes were then screened out. Cell apoptosis was detected by electron microscopy, in site cell apoptosis detection kit, DNA agarose gel electrophoresis and flow cytometry (FCM). RESULTS: Among the 82 genes which expressed differently after treatment with arsenic trioxide, 34 genes were up-regulated, while 48 genes down-regulated. It was detected that 15 micro mol/L As(2)O(3) can definitively induce HL60 cells to go apoptosis by FCM. Rate of apoptotic HL60 cells in control group is 1.7%, and As(2)O(3) group is 26.1%. CONCLUSION: Cyclin B1, proliferating cell nuclear antigen (PCNA), insulin like growth factor binding protein (IGFBP) et al may play an important role in HL60 cell apoptosis induced by arsenic trioxide.

[The significance of cyclin E expression and cyclin E-dependent kinase inhibitor in adult acute leukemia].

OBJECTIVE: To evaluate the function and clinical value of cyclin E and p27 in adult acute leukemia (AL). METHODS: The protein expression of cyclin E, p27, p170 and cell cycle distribution was measured in the bone marrow samples of 60 adult patients with AL and 17 controls with flow cytometric analysis. The mRNA levels of cyclin E, p27, MRP in 46 cases of AL and 17 controls were examined with reverse transcription-polymerase chain reaction method. RESULTS: The expression of cyclin E in AL patients was higher than that in controls (P < 0.01), so was the expression of p27 (P < 0.05). However in G(2)/M phase the expression rate of cyclin E in AL patients was not so apparently different with that in control (P < 0.05) so was that of p27 (P > 0.05). There was a positive correlation between the protein expression of cyclin E and p27. The remission rate (44.8%) in patients with over expression of cyclin E was lower than that (77.4%) in patients with normal expression of cyclin E (P < 0.01). The relapse rate (92.3%) in over expression group was higher than that (41.7%) in normal expression group (P = 0.003). No significant difference in remission rate or relapse rate was found between p27 over expression group and normal expression group (P > 0.50, P = 0.89). CONCLUSION: Over expression of cyclin E may implicate occurrence and progression of AL. Over expression of cyclin E is an important influential factor for remission rate and relapse rate in AL patients.

[The relationship between cyclin B1 and multidrug resistance in adult patients with acute leukemia].

OBJECTIVE: To investigate the relation between the expression of cyclin B1 and multidrug resistance in adult patients with acute leukemia. METHODS: The proteins expression of cyclin B1, p170 was measured with flow cytometric analysis in 85 adult de novo acute leukemia patients (AL) and 17 normal control (NC). The expression of cyclin B1, multidrug resistance gene (mdr-1), topoisomerase IIalpha, beta (TOPOIIalpha, beta) and bcl-2 mRNA in these patients was measured with semi-quantify reverse transcription polymerase chain reaction (RT-PCR). RESULTS: (1) The expression of cyclin B1 protein (M = 12.3%) and mRNA (M = 0.217) in the treatment resistance group was significantly lower than the sensitive group cyclin B1 protein (M = 22.7%) and mRNA (M = 0.563) (P < 0.05), so was the mRNA of TOPOIIalpha (M = 0.236), TOPOIIbeta (M = 0.328) than the sensitive group TOPOIIalpha (M = 0.514), TOPOIIbeta (M = 0.635) (P < 0.01). Cyclin B1 protein expression was lower than 5% and there were no expression of cyclin B1, TOPOIIalpha, mdr-1 mRNA in the NC group under the same condition. (2) The p170 protein (M = 14.3%) and mdr-1 (M = 1.071), bcl-2 (M = 0.941) mRNA expression in the resistant group was significant higher than the sensitive group p170 protein (M = 3.6%) and mdr-1 (M = 0.094), bcl-2 (M = 0.153) (P < 0.01). (3) The expression of cyclin B1 protein and TOPOIIalpha, TOPOIIbeta mRNA was positive correlated (r(TOPOIIalpha) = 0.472, P < 0.01; r(TOPOIIbeta) = 0.683, P < 0.01), so was the cyclin B1 mRNA (r(TOPOIIalpha) = 0.319, P < 0.05; r(TOPOIIbeta) = 0.527, P < 0.05). (4) There was no correlation between cyclin B1 and p170, mdr-1, bcl-2. (5) By Binary logistic forward conditional analysis we concluded that cyclin B1 correlated with atypital mutidrug resistance. CONCLUSIONS: Low expression of cyclin B1 might be a unfavorable prognostic factor for patients with AL and measurement of both cyclin B1 and TOPOIIalpha, TOPOIIbeta gene expression would predict drug resistance in adult acute leukemia patients.

[Expression of cyclin A in adult patients with acute leukemia].

OBJECTIVE: To investigate the clinical significance of cyclin A expression in adult patients with acute leukemia (AL). METHODS: 4 ml bone marrow was extracted from 100 AL patients and 10 normal controls to isolate the mononuclear cells (MNCs). The cyclin A mRNA levels in these MNCs were measured by RT-PCR. The cyclin A protein level and cell cycle in 75 randomly selected AL patients and 10 normal controls were examined by flow cytometric analysis. RESULTS: (1) The distribution of cyclin A protein was normal in cell cycle in AL patients. The positive rates of cyclin A protein and mRNA were 66.7% and 59%, significantly higher than those in the normal controls (0 and 1.4% respectively, both P < 0.01). The median expression levels of cyclin A protein and mRNA of cyclin A in AL patients were 18.5% and 0.539 +/- 0.490 respectively, significantly higher than those in the normal controls (both P < 0.01). Sequence analysis showed a complete consistency between the positive segment of cyclin A and the objective gene in GeneBank. (2) The levels of cyclin A protein and mRNA were positively correlated with the cumulative percentages of cells in S and G(2)/M phases (P < 0.01). (3) The expression level of cyclin A protein in the recurrent acute lymphoblastic leukemia (ALL) group was 15.4%, significantly lower than those of de novo group (29.5%, t = 14.418, P = 0.022). (4) The complete remission (CR) rates in the AL patients with high expression levels of cyclin A protein and mRNA group were 87.9% and 85.7% respectively, significantly higher than those in the AL patients with low expression levels (38.2% and 53.5% respectively, both P < 0.01). Multivariate regression analysis showed that cyclin A was one of the influencing factors of CR rate of AL patients (P < 0.05). CONCLUSION: Cyclin A expression contributes to the high proliferative activity in leukemia cells. The abnormal expression of cyclin A might be a prognostic marker of CR rate in AL patients.

[Effects of FPIL6/IL2 on the differentiation and proliferation of lymphoid in the ML-IC assay].

OBJECTIVE: To develop an in vitro assay that allows the culture and identification of a single human bone marrow progenitor closely related to hematopoietic stem cell, which is more primitive than LTC-IC, and to find an efficient culture conditions for NK-IC expansion. METHODS: Fusion protein IL6/IL2 was reconstructed and expressed in E. coli DH5 alpha. ML-IC was determined by watching if the single cell can give rise to secondary progenitors with both LTC-IC and NK-IC characteristics. LTC-IC frequency was determined by the CFC clonogenic methylcellulose assay. NK-IC frequency was determined by phenotyping CD56 positive NK cells. The effect of FPIL6/IL2 on the expansion of NK-IC was examined by comparing the colony number of NK cells before and after the culture. RESULTS: After the initial 4-week expansion culture, we showed that (25.75 +/- 5.68)% of freshly sorted Lin-/34+/DRdim cells were able to generate functional NK-IC in one or more secondary FPIL6/IL2 cultures, whereas (6.81 +/- 1.97)% in the control. A total of 102 NK-IC cells were present when were cultured for 6-7 weeks in FPIL6/IL2 expansion medium, which was much higher than the 33 NK-IC cells in the control. CONCLUSION: ML-IC assay will prove useful to assess a very primitive hematopoietic cell with multilineage generative capacity. FPIL6/IL2 is capable of initiating and promoting NK-IC expansion greatly in ex vivo cultures in terms of net-conservation and net proliferation.

[Characteristics of tenocyte adhesion to biologically-modified surface of polymer].

In this study we examined the in vitro characteristics of tenocyte adhesion to biologically-modified surface of polymer. Polylactic-co-glycolic acid (PLGA) 85/15 films were prepared by a solvent-casting technique. Each film was adhered onto the bottom of a chamber. The film was precoated with poly-D-lysine (PDL), and then coated with serum-free F12 medium containing various concentrations of fibronectin (FN), type I collagen (CN I), and insulin-like growth factor1 (IGF-1). The monoclonal antibodies (to FN and to CN I) with various dilutions were used to inhibit attachment of tenocytes to surface precoated with FN or CN I. Human embryonic tendon cells (HETCs) and transformed human embryonic tendon cells (THETCs) were used as the seeding cells. The system used for the measurement of adhesion force was the micropipette aspiration experiment system. The micropipette was manipulated to aspirate a small portion of the tenocyte body by using a small aspiration pressure. Then the pipette was pulled away from the adhesion area by micromanipulation. The minimum force required to detach the tenocyte from the substrate was defined as the adhesion force. The results showed that modification of FN or CN I by precoating significantly enhanced attachment of tenocytes to surface of polymer (P < 0.05). As antibodies to FN or CN I were added to a polymer film precoated with FN or CN I, the adhesion force decreased significantly (P < 0.05). We concluded that the specific adhesion forces of tenocytes to extracellular matrix adhesion proteins (FN and CN I) had coordinated action and showed good dependence on their precoating concentrations, and were inhibited by the antibodies to these adhesion proteins. Films precoated with IGF-1 strongly accelerated the adhesion of tenocytes to polymer. These results indicate that the specific adhesion of tenocytes to polymer can be promoted by coating extracellular matrix adhesive proteins and insulin-like growth factor1. It is of great importance to construct tissue-engineered tendon.