[Analysis of prognostic factors of extranodal NK/T-cell lymphoma treated with pegaspargase/L-asparaginase: a multicenter retrospective study].

Objective: To explore the prognostic factors of extracellular NK/T cell lymphoma (ENKTL) treated with pegaspargase/L-asparaginase. Methods: The clinical data of 656 ENKTL patients diagnosed at 11 medical centers in the Huaihai Lymphoma Working Group from March 2014 to April 2021 were retrospectively analyzed. The patients were randomly divided into two groups: a training set (460 cases) and a validation set (196 cases) at 7∶3, and the prognostic factors of the patients were analyzed. A prognostic scoring system was established, and the predictive performance of different models was compared. Results: Patients' median age was 46 (34, 57) years, with 456 males (69.5% ) and 561 nasal involvement (85.5% ). 203 patients (30.9% ) received a chemotherapy regimen based on L-asparaginase combined with anthracyclines, and the 5-year overall survival rate of patients treated with P-GEMOX regimen (pegaspargase+gemcitabine+oxaliplatin) was better than those treated with SMILE regimen (methotrexate+dexamethasone+cyclophosphamide+L-asparaginase+etoposide) (85.9% vs 63.8% ; P=0.004). The results of multivariate analysis showed that gender, CA stage, the Eastern Cooperative Oncology Group performance status (ECOG PS) score, HGB, and EB virus DNA were independent influencing factors for the prognosis of ENKTL patients (P<0.05). In this study, the predictive performance of the prognostic factors is superior to the international prognostic index, Korean prognostic index, and prognostic index of natural killer lymphoma. Conclusion: Gender, CA stage, ECOG PS score, HGB, and EB virus DNA are prognostic factors for ENKTL patients treated with pegaspargase/L-asparaginase.

[Mid-term outcome of deep layer repair with the long head of the biceps autograft bridging for Kim classification type â A delaminiated rotator cuff tear].

Objective: To investigate the mid-term clinical outcome of deep layer repair with the long head of the biceps autograft bridging for Kim classification type ⅠA delaminated rotator cuff tear. Methods: A follow-up study. The clinical data of 42 consecutive patients with Kim classification type ⅠA delaminated rotator cuff tear admitted to the First Affiliated Hospital of Jinan University from January 2018 to June 2019 were retrospectively included. All patients underwent shoulder arthroscopic surgery. During the operation, the autogenous long head of the biceps tendon was transferred to repair the deep layer of delaminated rotator cuff tear. The preoperative and postoperative (last follow-up) visual analogue scale (VAS) for pain, University of California Los Angeles (UCLA) score, Constant-Murley shoulder score, range of motion (ROM) of the shoulder and radiographic results were statistically analyzed. Results: A total of 42 patients were included in this study. There were 18 males and 24 females, with an average age of (64.5±15.2) years and a mean follow-up of (43.9±7.1) months. At the last follow-up, ROM of abduction increased from 80.8°±26.5° to 154.2°±14.3°, and ROM of external rotation increased from 18.2°±13.6° to 31.8°±7.8°; the VAS score of pain decreased from (5.5±1.3) points to (0.7±0.7) points, the UCLA score increased from (21.3±3.7) points to (29.9±2.1) points, and the Constant-Murley score increased from (45.4±10.0) points to (87.2±4.8) points; the differences were all statistically significant (all P<0.001). The X-ray films showed that there were no upward of the humeral head in all the patients. MRI results indicated that rotator cuff re-teared in one case (Sugaya classification type Ⅲ), and healed in other cases (Sugaya classification type Ⅰ-Ⅱ). No complications such as upper limb nerve injury was found in all cases. Conclusion: Deep layer repair with the long head of the biceps autograft bridging can significantly alleviate the pain and improve the function of patients with Kim classification type ⅠA delaminated rotator cuff tear, and the incidence of retear is low.

MiR-22-3p regulates the proliferation and invasion of Wilms' tumor cells by targeting AKT3.

OBJECTIVE: In this study, the regulatory mechanism of miR-22-3p/AKT3 in the development of Wilms' tumor (WT) was investigated. PATIENTS AND METHODS: Twenty-seven pairs of surgical tumor specimens and adjacent normal tissues were obtained from Jining No. 1 People's Hospital. The expression level of miR-22-3p in WT tissues and cell lines was measured by quantitative RT-PCR. MTT and transwell assays were performed to analyze cell proliferation and invasion in WT. The relationship between miR-22-3p and AKT3 was verified by a Dual-Luciferase assay. The protein expression of AKT3 was evaluated by Western blotting analysis. RESULTS: MiR-22-3p was downregulated and AKT3 was upregulated in WT. Functionally, overexpression of miR-22-3p inhibited cell proliferation and invasion in WT. Moreover, miR-22-3p directly targets AKT3. The knockdown of AKT3 suppressed cell proliferation and invasion in WT. In addition, upregulation of AKT3 restored the tumor suppressive effect of miR-22-3p in WT. CONCLUSIONS: MiR-22-3p inhibits the proliferation and invasion of WT cells by downregulating AKT3, indicating that miR-22-3p may be developed as a new biomarker for the diagnosis of WT.

H9N2 influenza virus isolated from minks has enhanced virulence in mice.

H9N2 is one of the major subtypes of influenza virus circulating in poultry in China, which has a wide host range from bird to mammals. Two H9N2 viruses were isolated from one mink farm in 2014. Phylogenetic analysis showed that internal genes of the H9N2 viruses have close relationship with those of H7N9 viruses. Interestingly, two H9N2 were separated in phylogenetic trees, indicating that they are introduced to this mink farm in two independent events. And further mice studies showed that one H9N2 caused obvious weight loss and 20% mortality in infected mice, while another virus did not cause any clinical sign in mice infected at the same dose. Genetic analysis indicated that the virulent H9N2 contain a natural mutation at 701N in PB2 protein, which was reported to contribute to mammalian adaptation. However, such substitution is absent in the H9N2 avirulent to mice. Circulation of H9N2 in mink may drive the virus to adapt mammals; continual surveillance of influenza virus in mink was warranted.

[Application of pegylated recombinant human granulocyte colony-stimulating factor to prevent chemotherapy-induced neutropenia in patients with lymphoma: a prospective, multicenter, open-label clinical trial].

Objective: To evaluate the efficacy and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in prophylaxis neutropenia after chemotherapy in patients with lymphoma. Methods: This was a multicenter, single arm, open, phase Ⅳ clinical trial. Included 410 patients with lymphoma received multiple cycles of chemotherapy and PEG-rhG-CSF was administrated as prophylactic. The primary endpoint was the incidence of Ⅲ/Ⅳ grade neutropenia and febrile neutropenia (FN) after each chemotherapy cycle. Meanwhile the rate of antibiotics application during the whole period of chemotherapy was observed. Results: ①Among the 410 patients, 8 cases (1.95%) were contrary to the selected criteria, 35 cases (8.54%) lost, 19 cases (4.63%) experienced adverse events, 12 cases (2.93%) were eligible for the termination criteria, 15 cases (3.66%) develpoed disease progression or recurrence, thus the rest 321 cases (78.29%) were into the Per Protocol Set. ②During the first to fourth treatment cycles, the incidences of grade Ⅳ neutropenia after prophylactic use of PEG-rhG-CSF were 19.14% (49/256) , 12.5% (32/256) , 12.18% (24/197) , 13.61% (20/147) , respectively. The incidences of FN were 3.52% (9/256) , 0.39% (1/256) , 2.54% (5/197) , 2.04% (3/147) , respectively. After secondary prophylactic use of PEG-rhG-CSF, the incidences of Ⅳ grade neutropenia decreased from 61.54% (40/65) in the screening cycle to 16.92% (11/65) , 18.46% (12/65) and 20.75% (11/53) in 1-3 cycles, respectively. The incidences of FN decreased from 16.92% (11/65) in the screening cycle to 1.54% (1/65) , 4.62% (3/65) , 3.77% (2/53) in 1-3 cycles, respectively. ③The proportion of patients who received antibiotic therapy during the whole period of chemotherapy was 34.39% (141/410) . ④The incidence of adverse events associated with PEG-rhG-CSF was 4.63% (19/410) . The most common adverse events were bone pain[3.90% (16/410) ], fatigue (0.49%) and fever (0.24%) . Conclusion: During the chemotherapy in patients with lymphoma, the prophylactic use of PEG-rhG-CSF could effectively reduce the incidences of grade Ⅲ/Ⅳ neutropenia and FN, which ensures that patients with lymphoma receive standard-dose chemotherapy to improve its cure rate.

Sequence variation in two mitochondrial DNA regions and internal transcribed spacer among isolates of the nematode Oesophagostomum asperum originating from goats in Hunan Province, China.

The present study examined sequence variability in two mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1), and internal transcribed spacer (ITS) of nuclear ribosomal DNA (rDNA) among Oesophagostomum asperum isolates from goats in Hunan Province, China. A portion of the cox1 (pcox1), nad1 (pnad1) genes and the ITS (ITS1+5.8S rDNA+ITS2) rDNA were amplified by polymerase chain reaction (PCR) separately from adult O. asperum individuals and the representative amplicons were subjected to sequencing from both directions. The lengths of pcox1, pnad1 and ITS rDNA were 366 bp, 681 bp and 785 bp, respectively. The A+T contents of gene sequences were 71.5-72% for pcox1, 73.7-74.2% for pnad1 and 58-58.8% for ITS rDNA. Intra-specific sequence variations within O. asperum were 0-1.6% for pcox1, 0-1.9% for pnad1 and 0-1.7% for ITS rDNA, while inter-specific sequence differences among members of the genus Oesophagostomum were significantly higher, being 11.1-12.5%, 13.3-17.7% and 8.5-18.6% for pcox1, pnad1 and ITS rDNA, respectively. Phylogenetic analyses using combined sequences of pcox1 and pnad1, with three different computational algorithms (Bayesian inference, maximum likelihood and maximum parsimony), revealed distinct groups with high statistical support. These findings demonstrated the existence of intra-specific variation in mtDNA and rDNA sequences among O. asperum isolates from goats in Hunan Province, China, and have implications for studying molecular epidemiology and population genetics of O. asperum.

Metabolomics for in situ environmental monitoring of surface waters impacted by contaminants from both point and nonpoint sources.

We investigated the efficacy of metabolomics for field-monitoring of fish exposed to wastewater treatment plant (WWTP) effluents and nonpoint sources of chemical contamination. Lab-reared male fathead minnows (Pimephales promelas, FHM) were held in mobile monitoring units and exposed on-location to surface waters upstream and downstream of the effluent point source, as well as to the actual effluent at three different WWTP sites in Minnesota. After four days of exposure, livers were collected, extracted, and analyzed by (1)H NMR spectroscopy and GC-MS to characterize responses of the hepatic metabolome. Multivariate statistical analysis revealed distinct metabolite profile changes in response to effluent exposure from each of the three WWTPs. Differences among locations (i.e., upstream, downstream, and effluent) within each of the three sites were also identified. These observed differences comport with land-use and WWTP characteristics at the study sites. For example, at one of the sites, the metabolomic analyses suggested a positive interactive response from exposure to WWTP effluent and nearby nonpoint (likely agricultural related) contamination. These findings demonstrate the utility of metabolomics as a field-based technique for monitoring the exposure of fish to impacted surface waters.

Field-based approach for assessing the impact of treated pulp and paper mill effluent on endogenous metabolites of fathead minnows (Pimephales promelas).

A field-based metabolomic study was conducted during a shutdown of a pulp and paper mill (PPM) to assess the impacts of treated PPM effluent on endogenous polar metabolites in fathead minnow (FHM; Pimephales promelas) livers. Caged male and female FHMs were deployed at a Great Lakes area of concern during multiple periods (pre-, during, and post-shutdown) near the outflow for a wastewater treatment plant. Influent to this plant is typically 40% PPM effluent by volume. Additional FHMs were exposed to reference lake water under laboratory conditions. A bioassay using T47D-KBluc cells showed that estrogenic activity of receiving water near the outflow declined by 46% during the shutdown. We then used (1)H NMR spectroscopy and principal component analysis to profile abundances of hepatic endogenous metabolites for FHMs. Profiles for males deployed pre-shutdown in receiving water were significantly different from those for laboratory-control males. Profiles were not significantly different for males deployed during the shutdown, but they were significant again for those deployed post-shutdown. Impacts of treated effluent from this PPM were sex-specific, as differences among profiles of females were largely nonsignificant. Thus, we demonstrate the potential utility of field-based metabolomics for performing biologically based exposure monitoring and evaluating remediation efforts occurring throughout the Great Lakes and other ecosystems.

Notch1 overexpression promotes cell growth and tumor angiogenesis in myeloma.

Patients with multiple myeloma (MM) have increased bone marrow angiogenesis, but the angiogenic properties of myeloma cells and the mechanism of MM-induced angiogenesis have not been completely clarified. Notch1 signal has been identified as a critical factor in the regulation of vessel formation. However, the role of Notch1 in the angiogenesis of MM is unclear. We constitutively overexpressed active Notch1 in RPMI8226 cells to explore the effect of Notch1 signaling on cell growth and tumor angiogenesis in vivo and in vitro. We found that Notch1 overexpression promoted myeloma cells growth and increased drug resistance. Moreover, vascular endothelial growth factor (VEGF) expression was increased. Finally, our in vitro results were supported by the in vivo finding in human myeloma xenograft Nonobese diabetic/severe combined immunodeficient (NOD/SCID) models. Notch1 overexpression in MM cells resulted in up-regulation of VEGF expression, promotion of tumor growth, and increased microvessel density (MVD). Our study suggests that Notch1-induced angiogenesis is partly due to activation of VEGF pathway.

Metabolite profiling and a transcriptional activation assay provide direct evidence of androgen receptor antagonism by bisphenol A in fish.

Widespread environmental contamination by bisphenol A (BPA) has created the need to fully define its potential toxic mechanisms of action (MOA) to properly assess human health and ecological risks from exposure. Although long recognized as an estrogen receptor (ER) agonist, some data suggest that BPA may also behave as an androgen receptor (AR) antagonist. However, direct evidence of this activity is deficient. To address this knowledge gap, we employed a metabolomic approach using in vivo exposures of fathead minnows (FHM; Pimephales promelas ) to BPA either alone or in a binary mixture with 17ß-trenbolone (TB), a strong AR agonist. Changes in liver metabolite profiles in female FHM in response to these exposures were determined using high resolution (1)H NMR spectroscopy and multivariate and univariate statistics. Using this approach, we observed clear evidence of the ability of BPA to mitigate the impact of TB, consistent with an antiandrogenic MOA. In addition, a transcriptional activation assay with the FHM AR was used to confirm the AR antagonistic activity of BPA in vitro. The results of these in vivo and in vitro analyses provide strong and direct evidence for ascribing an antiandrogenic MOA to BPA in vertebrates.

A pixel selection rule based on the number of different-phase neighbours for the simulated annealing reconstruction of sandstone microstructure.

Sandstone reservoir is one of the main types of oil and gas reservoirs in China. It has porous microstructure, which directly affects the transport properties of a sandstone. Hence, the study of porous microstructure is important to the exploration and exploitation of oil and gas. Three-dimensional microstructure of a sandstone can be reconstructed using the simulated annealing method based on statistical properties of its two-dimensional micrograph. The aim of reconstruction is to minimize the discrepancy between the statistical properties of the reconstructed microstructure and those of the two-dimensional image. To accelerate the rate of convergence, we proposed a different-phase neighbours (DPNs)-based pixel selection rule to replace the random pixel selection rule of the simulated annealing reconstruction. In this rule, pixels with the largest number of DPNs have the largest selection probability. The selection probabilities of other pixels are proportional to their DPNs. Microstructure reconstructed with the DPNs-based rule is compared with those with the random selection rule and two other biased pixel selection rules. The DPNs-based rule is the most effective in enhancing convergence. Permeability of the microstructure reconstructed with the DPNs-based rule is estimated by the Kozeny-Carman formula and is in good agreement with the one reconstructed with the random pixel selection rule.

Investigating compensation and recovery of fathead minnow (Pimephales promelas) exposed to 17alpha-ethynylestradiol with metabolite profiling.

1H NMR spectroscopy was used to profile metabolite changes in the livers of fathead minnows (Pimephales promelas) exposed to the synthetic estrogen 17alpha-ethynylestradiol (EE2) via a continuous flow water exposure. Fish were exposed to either 10 or 100 ng EE2/L for 8 days, followed by an 8 day depuration phase. Livers were collected after days 1, 4, and 8 of the exposure, and at the end of the depuration phase. Analysis of polar extracts of the liver revealed a greater impact of EE2 on males than females, with metabolite profiles of the former assuming similarities with those of the females (i.e., feminization) early in the exposure. Biochemical effects observed in the males included changes in metabolites relating to energetics (e.g., glycogen, glucose, and lactate) and liver toxicity (creatine and bile acids). In addition, amino acids associated with vitellogenin (VTG) synthesis increased in livers of EE2-exposed males, a finding consistent with increased plasma concentrations of the lipoprotein in the fish. Using partial least-squares discriminant analysis (PLS-DA), the response trajectories of the males at both exposure concentrations were compared. This revealed an apparent ability of the fish to compensate for the presence of EE2 later in the exposure, and to partially recover from its effects after the chemical was removed.

NMR analysis of male fathead minnow urinary metabolites: a potential approach for studying impacts of chemical exposures.

The potential for profiling metabolites in urine from male fathead minnows (Pimephales promelas) to assess chemical exposures was explored using nuclear magnetic resonance (NMR) spectroscopy. Both one-dimensional (1D) and two-dimensional (2D) NMR spectroscopy was used for the assignment of metabolites in urine from unexposed fish. Because fathead minnow urine is dilute, we lyophilized these samples prior to analysis. Furthermore, 1D 1H NMR spectra of unlyophilized urine from unexposed male fathead minnow and Sprague-Dawley rat were acquired to qualitatively compare rat and fish metabolite profiles and to provide an estimate of the total urinary metabolite pool concentration difference. As a small proof-of-concept study, lyophilized urine samples from male fathead minnows exposed to three different concentrations of the antiandrogen vinclozolin were analyzed by 1D 1H NMR to assess exposure-induced changes. Through a combination of principal components analysis (PCA) and measurements of 1H NMR peak intensities, several metabolites were identified as changing with statistical significance in response to exposure. Among those changes occurring in response to exposure to the highest concentration (450 microg/L) of vinclozolin were large increases in taurine, lactate, acetate, and formate. These increases coincided with a marked decrease in hippurate, a combination potentially indicative of hepatotoxicity. The results of these investigations clearly demonstrate the potential utility of an NMR-based approach for assessing chemical exposures in male fathead minnow, using urine collected from individual fish.

Anatomically discrete functional effects of adenoviral clostridial light chain gene-based synaptic inhibition in the midbrain.

The gene for the Light Chain fragment of Tetanus Toxin (LC) induces synaptic inhibition by preventing the release of synaptic vesicles. The present experiment applied this approach within the rat midbrain in order to demonstrate that LC gene expression can achieve functionally and anatomically discrete effects within a sensitive brain structure. The deep layers of the superior colliculus/deep mesencephalic nucleus (dSC/DpMe) that are located in the rostral midbrain has been implicated in fear-induced increase of the acoustic startle reflex (fear potentiated startle) but exists in close proximity to neural structures important for a variety of critical functions. The dSC/DpMe of adult rats was injected bilaterally with adenoviral vectors for LC, green fluorescent protein, or vehicle. Synaptobrevin was depleted in brain regions of adenoviral LC expression. LC gene expression in the dSC/DpMe inhibited the increase in startle amplitude seen with the control viral infection, and blocked context-dependent potentiation of startle induced by fear conditioning. Although LC gene expression reduced the absolute amount of cue-specific fear potentiated startle, it did not decrease percent potentiated startle to a cue, nor did it reduce fear-induced contextual freezing, nonspecific locomotor activity, or general health, indicating that its effects were functionally and anatomically specific. Thus, vector-driven LC expression inhibits the function of deep brain nuclei without altering the function of surrounding structures supporting its application to therapeutic neuromodulation.

Adenoviral clostridial light chain gene-based synaptic inhibition through neuronal synaptobrevin elimination.

Clostridial neurotoxins have assumed increasing importance in clinical application. The toxin's light chain component (LC) inhibits synaptic transmission by digesting vesicle-docking proteins without directly altering neuronal health. To study the properties of LC gene expression in the nervous system, an adenoviral vector containing the LC of tetanus toxin (AdLC) was constructed. LC expressed in differentiated neuronal PC12 cells was shown to induce time- and concentration-dependent digestion of mouse brain synaptobrevin in vitro as compared to control transgene products. LC gene expression in the rat lumbar spinal cord disrupted hindlimb sensorimotor function in comparison to control vectors as measured by the Basso-Beattie-Bresnahan (BBB) scale (P<0.001) and rotarod assay (P<0.003). Evoked electromyography (EMG) showed increased stimulus threshold and decreased response current amplitude in LC gene-transferred rats. At the peak of functional impairment, neither neuronal TUNEL staining nor reduced motor neuron density could be detected. Spontaneous functional recovery was observed to parallel the cessation of LC gene expression. These results suggest that light chain gene delivery within the nervous system may provide a nondestructive means for focused neural inhibition to treat a variety of disorders related to excessive synaptic activity, and prove useful for the study of neural circuitry.

A new complex variant t(4;15;17) in acute promyelocytic leukemia: fluorescence in situ hybridization confirmation and literature review.

We report a 37-year-old male with acute promyelocytic leukemia (APL) harboring a complex translocation (4;15;17). Karyotypic analysis with R-banding of bone marrow cells revealed 46,XY,t(4;15;17)(q21;q22;q21). Fluorescence in situ hybridization analysis using painting probes for chromosomes 4, 15 and 17 and reverse transcriptase polymerase chain reaction analysis revealed three derivative chromosomes: der(4)t(4;15)(q21;q22), der(15)t(4;15;17)(q21;q22;q21), and del(17)(q21q22). This is the third report of such a translocation and the first confirmed by molecular methods. Considering reported similar cases, it is possible that 4q21 is a nonrandom breakpoint in APL with complex translocations and the gene involved in 4q21 should be investigated.

Determination of perchlorate in tobacco plants and tobacco products.

Previous field and laboratory studies with vascular plants have shown that perchlorate is transported from perchlorate fortified soils and is accumulated in the plant tissues and organs. This paper contains results of preliminary investigations on the occurrence of perchlorate in tobacco plants grown in soils amended with a fertilizer whose nitrogen content is derived from naturally occurring sodium nitrate (Chile saltpeter). Ion chromatography (IC) and capillary electrophoresis (CE) were used for quantitative analysis, while nuclear magnetic resonance (NMR) spectroscopy was used for qualitative analysis of perchlorate. Results show that perchlorate is accumulated by tobacco plants into the leaves from soils amended with fertilizers that contain perchlorate. Also, perchlorate can persist over an extended period of time and under a variety of industrial processes as shown by its presence in off-the-shelf tobacco products including cigarettes, cigars, and pouch and plug chewing tobaccos in concentrations ranging from nd to 60.4 +/- 0.8 mg/kg on a wet weight basis.

[Edge-detection and its application in medical image processing].

Edge-detection is all the time a major problem in the computer early vision, and it plays an important role in image processing. This paper reviews classical and new methods of edge-detection and discusses its application in medical image processing.

[A contour map segmentation for laser scanning confocal microscopic biomedical images].

We propose a contour map segmentation method for laser scanning confocal microscopic (LSCM) biomedical images. In this method, an image is first segmented into sub-regions by contour map. Then the sub-regions are merged from top to bottom or from bottom to top, according to a segmentation criterion of intensity homogeneity. The merging procedures continue until no region needs to be merged. The advantage of this method is that the objects with wide intensity ranges and different intensity homogeneity can be segmented properly. In this paper, the region segmentation by contour map is first addressed. Next a region merging according to a segmentation criterion of intensity homogeneity is presented, and then the design of a linear intensity homogeneity segmentation criterion is introduced. Two examples of image segmentation by this method are given at the end of this paper.