A high-affinity potassium transporter (MeHKT1) from cassava (Manihot esculenta) negatively regulates the response of transgenic Arabidopsis to salt stress.

BACKGROUND: High-affinity potassium transporters (HKTs) are crucial in facilitating potassium uptake by plants. Many types of HKTs confer salt tolerance to plants through regulating K+ and Na+ homeostasis under salinity stress. However, their specific functions in cassava (Manihot esculenta) remain unclear. RESULTS: Herein, an HKT gene (MeHKT1) was cloned from cassava, and its expression is triggered by exposure to salt stress. The expression of a plasma membrane-bound protein functions as transporter to rescue a low potassium (K+) sensitivity of yeast mutant strain, but the complementation of MeHKT1 is inhibited by NaCl treatment. Under low K+ stress, transgenic Arabidopsis with MeHKT1 exhibits improved growth due to increasing shoot K+ content. In contrast, transgenic Arabidopsis accumulates more Na+ under salt stress than wild-type (WT) plants. Nevertheless, the differences in K+ content between transgenic and WT plants are not significant. Additionally, Arabidopsis expressing MeHKT1 displayed a stronger salt-sensitive phenotype. CONCLUSION: These results suggest that under low K+ condition, MeHKT1 functions as a potassium transporter. In contrast, MeHKT1 mainly transports Na+ into cells under salt stress condition and negatively regulates the response of transgenic Arabidopsis to salt stress. Our results provide a reference for further research on the function of MeHKT1, and provide a basis for further application of MeHKT1 in cassava by molecular biological means.

Critical bloodstream infection caused by Chromobacterium violaceum: a case report in a 15-year-old male with sepsis-induced cardiogenic shock and purpura fulminans.

Chromobacterium violaceum (C. violaceum) is a gram-negative bacillus that is widespread in tropical and subtropical areas. Although C. violaceum rarely infects humans, it can cause critical illness with a mortality rate above 50%. Here, we report the successful treatment of a 15-year-old male who presented with bloodstream infection of C. violaceum along with sepsis, specific skin lesions, and liver abscesses. Cardiogenic shock induced by sepsis was reversed by venoarterial extracorporeal membrane oxygenation (VA ECMO). Moreover, C. violaceum-related purpura fulminans, which is reported herein for the first time, was ameliorated after treatment. This case report demonstrates the virulence of C. violaceum with the aim of raising clinical awareness of this disease.

Positive Regulatory Roles of Manihot esculenta HAK5 under K+ Deficiency or High Salt Stress.

HAK/KUP/KT family members have been identified as playing key roles in K+ uptake and salt tolerance in numerous higher plants. However, their functions in cassava (Manihot esculenta Cantz) remain unknown. In this study, a gene encoding for a high-affinity potassium transporter (MeHAK5) was isolated from cassava and its function was investigated. Subcellular localization analysis showed that MeHAK5 is a plasma membrane-localized transporter. RT-PCR and RT-qPCR indicated that MeHAK5 is predominantly expressed in cassava roots, where it is upregulated by low potassium or high salt; in particular, its highest expression levels separately increased by 2.2 and 2.9 times after 50 µM KCl and 150 mM NaCl treatments. When heterologously expressed in yeast, MeHAK5 mediated K+ uptake within the cells of the yeast strain CY162 and rescued the salt-sensitive phenotype of AXT3K yeast. MeHAK5 overexpression in transgenic Arabidopsis plants exhibited improved growth and increased shoot K+ content under low potassium conditions. Under salt stress, MeHAK5 transgenic Arabidopsis plants accumulated more K+ in the shoots and roots and had reduced Na+ content in the shoots. As a result, MeHAK5 transgenic Arabidopsis demonstrated a more salt-tolerant phenotype. These results suggest that MeHAK5 functions as a high-affinity K+ transporter under K+ starvation conditions, improving K+/Na+ homeostasis and thereby functioning as a positive regulator of salt stress tolerance in transgenic Arabidopsis. Therefore, MeHAK5 may be a suitable candidate gene for improving K+ utilization efficiency and salt tolerance.

Cribriform-morular Thyroid Carcinoma Arising in a Medulloblastoma Survivor: Two Metachronous Tumors Shared with the Activation of the Wnt Signaling Pathway.

Wnt signaling pathway activation is involved in the pathogenesis of a series of malignant tumors and is characterized by the nuclear accumulation of ß-catenin protein. The occurrence of two or more Wnt pathway-associated tumors in a single individual is uncommon and generally attributed to inherited cancer syndrome, especially familial adenomatous polyposis (FAP). Herein, we presented a rare case of a child who suffered from the occurrence of Wnt-activated medulloblastoma and cribriform-morular thyroid carcinoma (CMTC) within a 9-year interval. She had no history of FAP and harbored an unexpected somatic mutation of the APC gene in the CMTC tumor. The potential agents involved in the pathogenesis of the two molecular-linked tumors other than FAP were discussed in this report.

Effect of public square dancing combined with serotonin reuptake inhibitors on persistent postural-perceptual dizziness (PPPD) in middle-aged and older women.

BACKGROUND: Persistent postural-perceptual dizziness (PPPD) is a functional vestibular disorder that causes chronic dizziness and limits daily activities. Although pharmacology, vestibular rehabilitation, and cognitive behavioral therapy have been proposed to have some efficacy, they have certain limitations. Some patients with PPPD report that public square dancing can effectively relieve the symptoms of dizziness and instability, and their mood improves. OBJECTIVE: To evaluate the effects of combining public square dancing with serotonin reuptake inhibitors (SSRIs/SNRIs) on the subjective sensations of dizziness, balance enhancement, anxiety, and depressive symptom regulation in middle-aged and older women with PPPD. MATERIALS AND METHODS: In this trial, 124 patients diagnosed with PPPD were enrolled. Among them, 64 patients were randomly assigned to the experimental group (EG), where they received square dance training combined with serotonin reuptake inhibitors. The remaining 60 cases were randomly assigned to the control group (CG), where they received only serotonin reuptake inhibitors and did not participate in organized sports activities, allowing them freedom in their daily lives. Data from the Dizziness Handicap Inventory (DHI), Hospital Anxiety and Depression Scale (HADS), Active-specific Balance Confidence Scale (ABC), and Vestibular Disorder Activities of Daily Living Scale (VADL) were collected and compared at the beginning, 3 months, and 6 months of the trial to evaluate the effect of public square dancing on middle-aged and older women with PPPD. RESULTS: There were no significant differences between the EG and CG before the trial. Compared with baseline measures, DHI, HADS, ABC, and VADL scores improved as the experiment progressed, and the improvements were more pronounced in the EG. CONCLUSION: Public square dancing combined with serotonin reuptake inhibitors has a positive impact on the subjective sensations of dizziness, balance enhancement, anxiety, and depressive symptom regulation in middle-aged and older women with PPPD.

High bifurcation of the unilateral brachial artery: an unusual variation.

BACKGROUND: It is common to observe a high bifurcation of the brachial artery, with some variations being quite rare. MATERIALS AND METHODS: During routine dissection for medical teaching purposes, a unilateral high bifurcation of the brachial artery was noted on the left side of a female cadaver. RESULTS: On the left arm, two branches of different sizes originate from the axillary artery, one on the radial side and the other on the ulnar side. These branches run down the arm, crossing each other, eventually becoming the forearm's radial and ulnar arteries. We also observed branches such as the posterior circumflex humeral artery, anterior circumflex humeral artery, and deep brachial artery. CONCLUSIONS: This case report adds to the existing body of knowledge about the variation of the brachial artery, which is of anatomical and clinical value. We report this case to highlight the unique variation in the arterial anatomy of the upper limb.

Preventive effect of cluster nursing on pressure ulcers in orthopedic patients and predictive value of serum IL-6 and TNF-α for the occurrence of pressure ulcers.

OBJECTIVE: To determine the effect of cluster nursing on pressure ulcer prevention and comfort of orthopedic patients. METHODS: A total of 124 orthopedic inpatients admitted to the Seventh Affiliated Hospital of Sun Yat-sen University from July 2018 to June 2021 were retrospectively analyzed. Among them, 66 cases received cluster nursing who were assigned into the observation group and the other 58 cases received routine nursing and were assigned into the control group. The incidence of pressure ulcers, the degree of pressure the ulcer, quality of life-brief (QOL-BREF), self-rating anxiety scale (SAS), and self-rating depression scale (SDS) scores of the two groups at 7 days after surgery were compared, and the comfort score and nursing satisfaction of the two groups were evaluated and compared. The expression and predictive value of serum interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in patients with pressure ulcers before operation were analyzed via enzyme-linked immunosorbent assay (ELISA). The levels of IL-6 and TNF-α were compared between the two groups before and after nursing. RESULTS: After nursing, the observation group had significantly lower IL-6, TNF-α, SAS score, and SDS scores than the control group, and showed significantly higher QOL-BREF score, nursing satisfaction and comfort scores than the control group. In addition, the observation group showed a significantly lower incidence of pressure ulcers and a significantly lower severity level than the control group after nursing. Patients with pressure ulcers showed significantly higher serum IL-6 and TNF-α levels before surgery, and receiver operating characteristic curve (ROC) showed that IL-6 and TNF-α had certain value in forecasting the occurrence of pressure ulcers. CONCLUSION: Cluster nursing can substantially lower the incidence of pressure ulcers in hospitalized orthopedic patients and improve their comfort.

IgG4-related disease with kidney and lymph nodes involvement: a case-based review.

IgG4-related disease (IgG4-RD), a rare immune-mediated chronic fibro-inflammatory condition, has various initial symptoms, thus posing diagnostic and therapeutic challenges. Here, we report a case of IgG4-RD in a 35-year-old man with initial clinical symptoms of facial edema and recent onset of proteinuria. It took more than 1 year from the onset of clinical symptoms to diagnosis. Pathological examination of renal biopsy revealed significant renal interstitial lymphoid tissue hyperplasia simulating growth pattern of lymphoma. Immunohistochemical (IHC) staining results showed that CD4 + T lymphocyte hyperplasia was dominant. There was no significant deletion of CD2/CD3/CD5/CD7. No monoclone was detected in TCR gene rearrangement. IHC staining showed that the number of IgG4-positive cells was greater than 100/HPF. The ratio of IgG4/IgG was greater than 40%. Combined with clinically examinations, IgG4-related tubulointerstitial nephritis was considered. Further cervical lymph node biopsy results suggested IgG4-related lymphadenopathy. He received methylprednisolone 40 mg/day intravenously for 10 days, leading to normal results of laboratory tests and clinical manifestations. The patient had a good prognosis without recurrence during 14 months of follow-up. This case report can be used as a reference for early diagnosis and treatment of such patients in the future.

Primary extranodal soft tissue Lennert lymphoma (lymphoepithelioid variant of peripheral T-cell lymphoma, unspecified): a case report and review of the literature.

Lennert lymphoma (LeL) is a rare variant of peripheral T-cell lymphoma, not otherwise specified (PTCL/NOS) that is rich in epithelioid histiocytes. LeL may pose great diagnostic and therapeutic challenges to the pathologist and clinician. Primary extranodal soft tissue LeL is even rarer and has not been reported. Herein, we report a case of LeL arising from soft tissue.A 65-year-old male presented for evaluation of a painless mass in the subcutaneous soft tissue of the left forehead. There was no invasion of the bone and no ulceration on the surface of the skin. The surrounding skin was erythematous and swollen. Grossly, the tumor was gray-red and 30 mm × 20 mm × 10 mm in size.Microscopically, the demarcation between the lesion and surrounding tissues was unclear without a capsule. The tumor invaded the surrounding striated muscle and adipose tissue. The tumor had a diffuse proliferation of small-sized atypical lymphocytes and numerous large clusters of epithelioid histiocytes. Plasma cells, eosinophils, and Hodgkin-Reed-Sternberg (HRS) cells were not identified. Rare multinucleated histiocytes were noted, and well-formed granulomas were not present. Rare mitotic figures were noted, but no necrosis. The immunophenotypic features in this case were as follows: CD2+/CD3+/CD5low+/CD7+/CD4low+/ CD8+/CD30-/CD56- in neoplastic lymphocytes; CD163+/CD31+/CK(pan)- in epithelioid histiocytes; and CD20-/CD30-/TdT-/CD5-/ALK-/S-100-/CD1α-/CD21 + 23-/SSTR2- in neoplastic lymphocytes and epithelioid histiocytes. Epstein-Barr virus (EBV)-encoded RNA in situ hybridization (EBER-ISH) was negative. The Ki-67 index was elevated to 60%. PCR showed a polyclonal pattern for IgH and a monoclonal TCR γ-chain rearrangement.The final diagnosis was PTCL/NOS, lymphoepithelioid cell variant (LeL), which arose from soft tissue and had a rare double-positive CD4low+/CD8+ immunophenotype. The patient received four cycles of cyclophosphamide, doxorubicin liposomes, vincristine, and prednisone tablets (CHOP) and was followed for 20 months. Overall treatment efficacy was achieved without lymphadenopathy, and no other discomfort or illnesses were reported.

MeCIPK10 regulates the transition of the K+ transport activity of MeAKT2 between low- and high-affinity molds in cassava.

AKT1 is an inward-rectifying K+ channel that was originally thought to function only within a low-affinity K+ concentration range. However, the growth of an akt1 mutant of Arabidopsis was shown to be severely inhibited within a high-affinity range. This suggested that AKT1 may also be a high-affinity K+ transporter, but it remains unclear how the two modes of AKT1 coordinate to uptake K+. One gene (MeAKT2) encodes for a putatively inward-rectifying K+ channel and was isolated from cassava. Relative to other tissues, the MeAKT2 gene was expressed mainly in roots, and its transcriptional level was observed to be significantly increased under low-K+ conditions. Functional analyses were performed using a yeast expression system. When MeAKT2 was expressed alone in yeast cells, transgenic yeast could grow only in nutrient media supplied with >0.5 mM potassium. A yeast two-hybrid assay showed that both MeCIPK10 and MeCIPK12 clearly interacted with MeAKT2. Additionally, 0.05 mM K+ was sufficient for the growth of yeast cells co-expressing MeAKT2 with MeCIPK10, but also their co-expression significantly enhanced the growth capacity of yeast cells in the low range of K+ concentrations. Change in K+ uptake rate in co-transgenic yeast cells grown across a wide range of K+ concentrations showed that MeAKT2-mediated K+ uptake displayed a biphasic pattern, but also the switching from low-to high-affinity K+ uptake was regulated by CIPK10. This indicated that MeAKT2 functioned as a dual-affinity transporter to uptake K+ under both low- and high-affinity K+ conditions.

The chloroplast genome characteristics, comparative genomics and gene resource mining of Celtis sinensis (Persoon, 1805).

Celtis is a large genus in Cannabaceae family, with more than 70 species in the world. However, the intraspecific variabilities of morphological features make it difficult for some species to be distinguished based on their morphological characteristics. To supply the chloroplast (cp) genome resources of Celtis for species identification, the plastome of Celtis sinensis Persoon 1805 was newly sequenced and comparative genomics was analyzed. The chloroplast genome was 159,085 bp in length and had a quadripartite structure consisting of two inverted repeats (IRs) separated by a small single copy (SSC) and a large single copy (LSC) region. A total of 133 genes were annotated, including 88 protein-coding genes, eight rRNA genes, and 37 tRNA genes. Among the protein-coding genes, the frequency of the leucine codon is the highest and that of the cysteine codon is the lowest. Comparative genomic analysis showed that the IRS region was more conservative than the LSC and SSC regions, with most sequence variations located in the intergenic spacer rather than the protein-coding region. Moreover, sixteen highly divergent hotspots were identified. The ML phylogenetic tree showed that all involved Celtis species were clustered together, and the plastome reported in this paper has high enough resolution to distinguish C. sinensis (Pers.) from other Celtis plants. This study provides useful genetic resources for the identification of C. sinensis (Pers.) and is also of great significance for the phylogeny study of Celtis plants in the future.

Outcome of Extracorporeal Membrane Oxygenation Combined with Intraaortic Balloon Pump Hemodynamic Support during the Percutaneous Coronary Intervention Process for Patients with Cardiac Shock Complicating Acute Myocardial Infarction.

We evaluate the effect of extracorporeal membrane oxygenation combined with intraaortic balloon pump mechanical circulatory support for patients with cardiogenic shock complicating acute myocardial infarction during the PCI process. Extracorporeal membrane oxygenation combined with intraaortic balloon pump hemodynamic support during the percutaneous coronary intervention process for patients with cardiac shock complicating acute myocardial infarction might play a complementary role. Yet, evidence of application of both devices at the same time remains unclear. Patients with cardiogenic shock complicating myocardial infarction who underwent PCI in our hospital from January 2015 to January 2018 were screened. Those who were under hemodynamic support of extracorporeal membrane oxygenation combined with intraaortic balloon pump were enrolled as the ECMO&IABP group, and the patients only under support of intraaortic balloon pump were enrolled as the IABP group. The differences of clinical prognosis between the two groups were compared. A total of 39 patients were enrolled into the study: 10 were in the ECMO&IABP group and 29 in the IABP group. Compared with the IABP group, more patients were complicated with old myocardial infarction (5/10 vs. 2/29, p=0.002), more patients were diagnosed as non-ST elevated myocardial infarction (8/10 vs. 11/29, p=0.002) and left ventricular ejecting fraction was lower (41.1 ± 9.86 vs. 48.55 ± 8.86, p=0.03) in the ECMO&IABP group. Mechanical complications were higher in the ECMO&IABP group (5/10 vs. 5/29, p=0.048), The survive rate in the ECMO&IABP group is higher than that in the IABP group (90.00% vs. 47.83%, p=0.042) at one-year follow-up. Compared with only IABP, ECMO combined with IABP hemodynamic support during the PCI process for patients with cardiogenic shock complicating acute myocardial infarction enjoys better mortality outcome.

Genome-wide analysis and phosphorylation sites identification of the 14-3-3 gene family and functional characterization of MeGRF3 in cassava.

The biological functionality of many members of the 14-3-3 gene family is regulated via phosphorylation at multiple amino acid residues. The specific phosphorylation-mediated regulation of these proteins during cassava root tuberization, however, is not well understood. In this study, 15 different 14-3-3 genes (designated MeGRF1 - 15) were identified within the cassava genome. Based upon evolutionary conservation and structural analyses, these cassava 14-3-3 proteins were grouped into ε and non-ε clusters. We found these 15 MeGRF genes to be unevenly distributed across the eight cassava chromosomes. When comparing the expression of these genes during different developmental stages, we found that three of these genes (MeGRF9, 12 and 15) were overexpressed at all developmental stages at 75, 104, 135, 182 and 267 days post-planting relative to the fibrous root stage, whereas two (MeGRF5 and 7) were downregulated during these same points. In addition, the expression of most MeGRF genes changed significantly in the early and middle stages of root tuberization. This suggests that these different MeGRF genes likely play distinct regulatory roles during cassava root tuberization. Subsequently, 18 phosphorylated amino acid residues were detected on nine of these MeGRF proteins. A phosphomimetic mutation at serine-65 in MeGRF3 in Arabidopsis thaliana (Arabidopsis) slightly influenced starch metabolism in these plants, and significantly affected the role of MeGRF3 in salt stress responses. Together these results indicate that 14-3-3 genes play key roles in responses to abiotic stress and the regulation of starch metabolism, offering valuable insights into the functions of these genes in cassava.

Integrin ß4 promotes invasion and anoikis resistance of papillary thyroid carcinoma and is consistently overexpressed in lymphovascular tumor thrombus.

Although the majority of papillary thyroid cancers (PTC) are indolent, a subset of PTCs behaves aggressively due to extensive invasion and distant metastasis. Integrin ß4, a member of the integrin family, has been shown to enhance the progression in some malignancies; however, its role in PTC remains unclear. Here, we demonstrated that ß4 overexpression was associated with extrathyroid extension, lymph node metastasis, high TNM stage, and poor overall survival based on The Cancer Genome Atlas cohort. Immunohistochemistry showed that ß4 expression was significantly upregulated in the tumors with infiltrating growth pattern, as well as those with positive lymphovascular invasion. Moreover, ß4 was invariably overexpressed in the lymphovascular tumor thrombi, which has not been reported before. After shRNA-induced knockdown of ß4 in vitro, the migration, invasion and scratch repair ability of the tumor cells were significantly reduced. Furthermore, ß4 reduction decreased anchorage-independent growth and increased anoikis. The bioinformatics analysis revealed that approximately 70 pathways were significantly dysregulated in the high ß4 expression group. The MAPK pathway and propanoate metabolism were located in the network center of those pathways. Taken together, our results suggest that ß4 could promote the tumor's aggressiveness by enhancing invasion and antagonizing anoikis. The upregulated expression of ß4 in the tumor thrombi is intrinsically linked to its role in strengthening the anoikis resistance.

Characterization of inflammatory factor-induced changes in mesenchymal stem cell exosomes and sequencing analysis of exosomal microRNAs.

BACKGROUND: Treatments utilizing stems cells often require stem cells to be exposed to inflammatory environments, but the effects of such environments are unknown. AIM: To examine the effects of inflammatory cytokines on the morphology and quantity of mesenchymal stem cell exosomes (MSCs-exo) as well as the differential expression of microRNAs (miRNAs) in the exosomes. METHODS: MSCs were isolated from human umbilical tissue by enzymatic digestion. Exosomes were then collected after a 48-h incubation period in a serum-free medium with one of the following the inflammatory cytokines: None (control), vascular cell adhesion molecule-1 (VCAM-1), tumor necrosis factor (TNF) α, and interleukin (IL) 6. The morphology and quantity of each group of MSC exosomes were observed and measured. The miRNAs in MSCs-exo were sequenced. We compared the sequenced data with the miRBase and other non-coding databases in order to detect differentially expressed miRNAs and explore their target genes and regulatory mechanisms. In vitro tube formation assays and Western blot were performed in endothelial cells which were used to assess the angiogenic potential of MSCs-exo after inflammatory cytokine stimulation. RESULTS: MSCs-exo were numerous, small, and regularly shaped in the VCAM-1 group. TNFα stimulated MSCs to secrete larger and irregular exosomes. IL6 led to a reduced quantity of MSCs-exo. Compared to the control group, the TNFα and IL6 groups had more downregulated differentially expressed miRNAs, particularly angiogenesis-related miRNAs. The angiogenic potential of MSCs-exo declined after IL6 stimulation. CONCLUSION: TNFα and IL6 may influence the expression of miRNAs that down-regulate the PI3K-AKT, MAPK, and VEGF signaling pathways; particularly, IL6 significantly down-regulates the PI3K-AKT signaling pathway. Overall, inflammatory cytokines may lead to changes in exosomal miRNAs that abnormally impact cellular components, molecular function, and biological processes.

MicroRNA­944 targets vascular endothelial growth factor to inhibit cell proliferation and invasion in osteosarcoma.

Emerging evidence has demonstrated that the dysregulation of microRNA (miRNA/miR) serves a crucial role in the tumorigenesis and tumor development of osteosarcoma (OS), primarily by affecting various pathological behaviors. Therefore, better knowledge of miRNA in OS may provide novel insights into the pathogenesis of OS, and may facilitate the development of promising therapeutics for patients with this disease. MiRNA­944 is frequently dysregulated in human cancers. However, the expression levels, functions and underlying mechanisms of miR­944 in OS remain largely elusive. In the present study, reverse transcription­quantitative polymerase chain reaction (RT­qPCR) was performed to detect miR­944 expression in OS tissues and cell lines. The regulatory influence of miR­944 in OS proliferation and invasion was determined with MTT and Transwell invasion assays. In addition, the mechanisms underlying the action of miR­944 in OS cells were elucidated through a series of experiments, including bioinformatics analysis, luciferase reporter assay, RT­qPCR and western blot analysis. Spearman's correlation analysis was utilized to examine the relationship between miR­944 and VEGF expression levels, and rescue experiments were applied to further verify whether VEGF mediates the role of miR­944 in OS. The results demonstrated that miR­944 was downregulated in cancer tissues and cell lines. Furthermore, exogenous miR­944 expression inhibited cell proliferation and invasion in OS in vitro. Vascular endothelial growth factor (VEGF) was identified as a direct target of miR­944 in OS and was overexpressed in cancer tissues. VEGF expression was inversely correlated with miR­944 expression in cancer tissues. Rescue experiments demonstrated that overexpression of VEGF partially prevented the miR­944­induced inhibition of OS cell proliferation and invasion. These results suggested that miR­944 may serve a tumor suppressive role in OS by directly targeting VEGF. Therefore, miR­944 may be a promising target in the treatment of OS.

[Karyotype analysis of three Lonicera species growth in Sichuan].

OBJECTIVE: To provide the genetic reference for development and application of Lonicera species growth in Sichuan. METHODS: Shoot apices cells from Lonicera japonica, Lonicera japonica var. chinensis and Lonicera similis (cultivars) were used to make the chromosomal preparation. Their karyotypes were analyzed and the relevant parameters of chromosomes were measured by improved chromosome preparation technique. RESULTS: The chromosome numbers of three Lonicera species were 2n = 2X = 18; their chromosome length was 30.747, 33.231 and 36.948 microm; Their karyotype formula was 2n = 2X = 18 = 2m + 7sm, 2n = 2X = 18 = 8m + 8sm + 2st and 2n = 2X = 18 = 8sm + 10m; Their As. k was 64.013%, 64.380% and 61.949%; And their karyotypes belonged to 2B, 2B and 2A type, respectively. CONCLUSION: These three Lonicera species can be of the germplasm resources for widely cultivating since they have high degree genetic evolution.

[Induction on callus culture and regeneration of Orostachyis fimbriatae].

OBJECTIVE: To explore the effects of different hormonal combinations on induction, proliferation and differentiation of Orostachyis fimbriatae callus culture. METHODS: Aseptic seedling leaves were used as explants,the different concentrations of IAA,NAA, 6-BA and KT on induction proliferation of callus were optimized by orthogonal test to explore the optimum medium for differentiation of callus by tissue culture techniques. RESULTS: The best medium for induction was MS + IAA 1.0 mg/L + NAA 0.5 mg/L + KT 1.0 mg/L, and the best hormonal combination for proliferation was MS + IAA 0.5 mg/L + 6-BA 0.5 mg/I. + KT 1.0 mg/L. The best medium for differentiation was MS + IAA 0.1 mg/L + KT 2.0 mg/L, and 1/2MS + IAA 0.2 mg/L was the optimum medium for rooting culture. CONCLUSION: The system of regeneration of Orostachyis fimbriatae is establishd by tissue culture techniques in this study.

Immunohistochemistry in the diagnosis of papillary lesions of the breast.

AIMS: Many immunohistochemical markers have been studied for differentiating papillary lesions. However, their differentiating power has not been evaluated comprehensively. Additionally, assessment of some markers will require the difficult task of identifying different cell types. In the current study, we aimed to devise a simple papillary panel which can aid in diagnosis irrespective of architectural pattern and cell type differentiation. METHODS AND RESULTS: Immunohistochemical analysis of papillary lesions using myoepithelial markers [p63 and smooth muscle actin (SMA)], high molecular weight cytokeratins (HMWCKs: CK5, CK5/6, CK14 and 34ßE12), hormone receptors (ER and PR) and neuroendocrine markers (chromogranin and synaptophysin) was performed. Among them, neuroendocrine markers showed high specificity but low sensitivity. HMWCK specificity was better than that for myoepithelial markers. Homogeneous staining pattern for hormonal receptors rather than their percentage positivity was more effective in identifying malignant lesions. Negative staining for two or more of HMWCKs, namely CK5/6, CK14 and 34ßE12, achieved the best overall specificity and sensitivity of 87.8% and 94.1%, respectively, irrespective of the architecture. Their discriminatory power was validated with an independent cohort of core needle biopsies. CONCLUSIONS: A marker panel with HMWCKs could be used in differentiating papillary lesions irrespective of architectural pattern or cell type differentiation.

Increased SOX2 expression in less differentiated breast carcinomas and their lymph node metastases.

AIMS: SOX2 is a key regulatory gene in embryonic stem cells. Although it has been implicated in cancer progression, its role in breast carcinoma is poorly understood. MATERIALS AND METHODS: Fifty-seven ductal carcinomas in situ (DCIS), 552 invasive breast carcinomas and 107 corresponding metastatic lymph nodes were evaluated immunohistochemically for the expression of SOX2. Its correlation with clinicopathological features, other biomarker profiles and patients' outcomes were analysed. RESULTS: SOX2 was detected in 19.0% (105 of 552) of invasive breast carcinomas and 12.3% (seven of 57) of DCIS. Expression correlated with larger tumour size (P = 0.005) and higher grade (P = 0.002). It was associated negatively with ER (P = 0.015) and PR (P = 0.046) expression, but positively with Ki67 index (P = 0.013). Interestingly, it was also associated with neuroendocrine marker expression (synpatophysin and chromogranin/synaptophysin, P = 0.048 and 0.028, respectively). Expression appeared to be independent from that of common stem cell markers, namely CD44, CD24 and aldehyde dehydrogenase 1 (ALDH1). Furthermore, a higher rate of expression was observed in metastatic lymph nodes than in the corresponding primary tumours (P = 0.034). High SOX2 expression was correlated with poor disease-free survival (log-rank=9.489, P = 0.012) and was an independent prognostic factor (HR=2.918, P = 0.015) in patients with high nodal stages. CONCLUSIONS: In summary, SOX2 expression was related to adverse breast carcinoma profile and poor outcome in selected patient groups.