Photoregulatory protein kinases fine-tune plant photomorphogenesis by directing a bifunctional phospho-code on HY5 in Arabidopsis.

Photomorphogenesis is a light-dependent plant growth and development program. As the core regulator of photomorphogenesis, ELONGATED HYPOCOTYL 5 (HY5) is affected by dynamic changes in its transcriptional activity and protein stability; however, little is known about the mediators of these processes. Here, we identified PHOTOREGULATORY PROTEIN KINASE 1 (PPK1), which interacts with and phosphorylates HY5 in Arabidopsis, as one such mediator. The phosphorylation of HY5 by PPK1 is essential to establish high-affinity binding with B-BOX PROTEIN 24 (BBX24) and CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1), which inhibit the transcriptional activity and promote the degradation of HY5, respectively. As such, PPKs regulate not only the binding of HY5 to its target genes under light conditions but also HY5 degradation when plants are transferred from light to dark. Our data identify a PPK-mediated phospho-code on HY5 that integrates the molecular mechanisms underlying the regulation of HY5 to precisely control plant photomorphogenesis.

Anti-MDA5 antibody dermatomyositis-associated rapidly progressive interstitial lung disease patient complicated with mixed connective tissue disease: A case report.

Anti-MDA5 antibody dermatomyositis (DM) is a special type of myositis, which can potentially cause rapidly progressive interstitial lung disease (RP-ILD). Mixed connective tissue disease (MCTD) is a complex disease with different characteristics of autoimmune connective tissue disease, associated with ILD. Both are rare diseases, and few patients with both diseases have been reported. A 71-year-old woman complained of palpitations, with a 2 months history of rash around her hands, extensor surface of right elbow, and the nape of her neck. Subsequently, the patient had acute exacerbation of dyspnea and tachypnea. Anti-Ro52, U1 RNP and MDA5 antibodies were positive; the presenting evidence was suggestive of anti-MDA5+ DM-RP-ILD complicated with MCTD. Our patient deteriorated rapidly and had a fatal outcome, despite "triple therapy" for RP-ILD. This case illustrates that patients with coexisting anti-MDA5+ DM and MCTD have the former's typical clinical manifestations, and may develop ILD quickly rather than slowly as in MCTD, especially with the coexistence of anti-Ro52 antibodies.

Selective bronchial occlusion for the prevention of pneumothorax after transbronchial lung cryobiopsy in a pulmonary alveolar proteinosis patient: a case report.

The diagnosis of pulmonary alveolar proteinosis (PAP) is based on biopsies. Compared with other methods of taking biopsies, transbronchial lung cryobiopsy (TBLC) has a higher diagnostic rate and the likelihood of pneumothorax. Selective bronchial occlusion (SBO) is an effective technique for treating intractable pneumothorax. However, there are no data available about SBO for the prevention of pneumothorax after TBLC in a PAP patient. A 49-year-old man complained of recurrent cough and tachypnea, and his symptoms did not fully resolve until the diagnosis was confirmed, and he was treated with whole lung lavage. Our patient was ultimately diagnosed with PAP by TBLC but not multiple tests for the bronchoalveolar lavage fluid (BALF). The patient was discharged quickly after whole lung lavage due to the fact that he did not develop pneumothorax under SBO. This case illustrates that TBLC is a supplementary examination for PAP, especially for those in whom BALF results fail to confirm a diagnosis. Moreover, our report highlights that SBO is necessary to effectively prevent pneumothorax during and after multiple TBLCs in PAP patients.

The receptor kinase OsWAK11 monitors cell wall pectin changes to fine-tune brassinosteroid signaling and regulate cell elongation in rice.

Rates of plant cell elongation change with day-night alternation, reflecting differences in metabolism related to cell wall remodeling. Information from cell wall surveillance pathways must be integrated with growth regulation pathways to provide feedback regulation of cell wall modification; such feedback regulation is important to ensure sufficient strength and prevent rupture of the cell wall during growth. Several lines of evidence suggest that cell wall perturbations often influence phytohormone signaling, but the identity of the nexus between these two processes remained elusive. Here, we show that wall-associated kinase11 (OsWAK11) acts as a linker connecting cell wall pectin methyl-esterification changes and brassinosteroid (BR) signaling in rice. Our data show that OsWAK11 controls several important agronomical traits by regulating cell elongation in rice. OsWAK11 directly binds and phosphorylates the BR receptor OsBRI1 at residue Thr752, within a motif conserved across most monocot graminaceous crops, thus hindering OsBRI1 interaction with its co-receptor OsSERK1/OsBAK1 and inhibiting BR signaling. The extracellular domain of OsWAK11 shows a much stronger interaction toward methyl-esterified pectin as compared with de-methyl-esterified pectin. OsWAK11 is stabilized in light but is degraded in darkness, in a process triggered by changes in the ratio of methyl-esterified to de-methyl-esterified pectin, creating fluctuations in plant BR signaling in response to day and night alternation. We conclude that OsWAK11 is a cell wall monitor that regulates cell elongation rates to adapt to the environment from the outside in, which complements the well-established inside-out signaling pathway affecting cell elongation in plants.

[Result analysis of percutaneous core needle biopsy for bone tumors in upper limbs with pathological fracture].

OBJECTIVE: To analyze the results of percutaneous core needle biopsy for bone tumors in upper limbs with pathologic fracture and to find the possible factors that could impact the results. METHODS: The including criteria for this study was the patients who had received percutaneous core needle biopsy and definitive surgery, whose tumor was located at upper limb with pathologic fracture. From January 2015 to December 2019, seventy-seven patients were enrolled. There were 55 males and 22 females. The median age was 27 years old (range:5 to 88 years old). The tumor located at humerus in 67 cases, radius in 8 cases and ulna in 2 cases. If the pathologic diagnosis of core needle biopsy was the same with the definitive surgery, it was defined as "correct". If the pathologic diagnosis of biopsy for benign or malignant was right but the exact diagnostic name was not the same with definitive surgery, it was defined as "supportive". If the pathologic diagnosis of biopsy for benign or malignant was not correct, it was defined as "wrong". We retrospectively analyzed the accuracy and impact factors for core needle biopsy. RESULTS: The result was "correct" in 63 cases(81.8%), "supportive" in 14 cases(18.2%), and "wrong" in 0 cases. We analyzed the gender, age, location, fracture displacement, the destroyed type for bone tumor, soft tissue mass, fluid area in the tumor as the factors. The results showed the rate for "correct" was significantly higher when the tumor had soft tissue mass (P< 0.05) and lower when the fluid area existed inside the tumor (P<0.05). CONCLUSION: The accuracy of percutaneous core needle biopsy for upper limb bone tumor with pathologic is high and acceptable. The biopsy chosen the soft tissue mass area can increase the accuracy.

Mutual Regulation of Receptor-Like Kinase SIT1 and B'κ-PP2A Shapes the Early Response of Rice to Salt Stress.

The receptor-like kinase SIT1 acts as a sensor in rice (Oryza sativa) roots, relaying salt stress signals via elevated kinase activity to enhance salt sensitivity. Here, we demonstrate that Protein Phosphatase 2A (PP2A) regulatory subunit B'κ constrains SIT1 activity under salt stress. B'κ-PP2A deactivates SIT1 directly by dephosphorylating the kinase at Thr515/516, a salt-induced phosphorylation site in the activation loop that is essential for SIT1 activity. B'κ overexpression suppresses the salt sensitivity of rice plants expressing high levels of SIT1, thereby contributing to salt tolerance. B'κ functions in a SIT1 kinase-dependent manner. During early salt stress, activated SIT1 phosphorylates B'κ; this not only enhances its binding with SIT1, it also promotes B'κ protein accumulation via Ser502 phosphorylation. Consequently, by blocking SIT1 phosphorylation, B'κ inhibits and fine-tunes SIT1 activity to balance plant growth and stress adaptation.

Magnetic Resonance Imaging for the Assessment of Long Bone Tumors.

BACKGROUND: Wide resection margins of osseous tumors are associated with a low incidence of local recurrence, making accurate measurement of the intraosseous extent of primary malignant long bone tumors is crucial. We compared the intraosseous tumor extent assessed by magnetic resonance imaging (MRI) with the gross specimen to evaluate the accuracy of MRI. METHODS: A total of 255 patients with primary malignant tumors in the long bones were included. Using MRI, we defined the length of tumor as the distance from the articular surface to the boundary between abnormal and normal marrow signal. The extent of the abnormal intraosseous signal was measured on unenhanced T1-weighted (T1WI) magnetic resonance images after chemotherapy. All gross surgical specimens were sectioned, and tumor extent was measured. Wilcoxon signed-rank test was used to test the differences between MRI and gross specimen findings. Spearman's correlation analysis was used to test the correlation between groups. RESULTS: Median tumor length by gross specimen (112 mm; range, 45-300 mm) was longer than that by MRI (108 mm; range, 45-304 mm; Z = -6.916, P < 0.001). Of 255 images, tumor length was accurately represented on 27 T1WI magnetic resonance images, overestimated on 79 images, and underestimated on 149 images. The median difference between imaging and gross specimen measurements was 2.0 mm (range: 1.0-15.0 mm) for the 79 cases where tumor length was overestimated, and 5.0 mm (range: 1.0-18.0 mm) for the 149 cases where tumor length was underestimated. The Spearman correlation demonstrated a high correlation of tumor length on gross specimen with the tumor length on MRI (R = 0.99, P < 0.01). CONCLUSIONS: We conclude that preoperative MRI could be a useful method in determining intramedullary malignant bone tumor boundaries and may serve as an accepted assessment method of long bone tumors before limb-sparing surgery.

Transition from Tumor Tissue to Bone Marrow in Patients with Appendicular Osteosarcoma after Neoadjuvant Chemotherapy.

BACKGROUND:: Limb-salvage surgery is the standard procedure for the treatment of appendicular osteosarcoma. Precise resection is the trend in limb-salvage surgery. The aim of this study was to evaluate a large series of cases to identify the histological relationship between the tumor and marrow and determine the intramedullary transition type and width from the tumor to normal marrow in patients with osteosarcoma after neoadjuvant chemotherapy. METHODS:: One hundred and six osteosarcoma specimens were evaluated. The tissue specimens were sectioned through the coronal axis by an electronic saw. The tissue was immersed in formalin solution for fixation and subsequently decalcified. The interface between the tumor and normal bone marrow was grossly determined and submitted for microscopic evaluation to detect the relationship between the tumor and bone marrow and identify the transition type and width. All histological slides were examined by experienced orthopedic pathologists. RESULTS:: Histologically, the interface between the tumor and normal bone marrow was classified into two patterns: "clear" and "infiltrated." The clear pattern, characterized by a clear boundary between the tumor and marrow, was identified in sixty cases (56.6%). A subtype of the clear type, characterized by fibrous bands between the tumor and marrow, was found in 13 cases (12.3%). The infiltrated pattern, characterized by a boundary with tumor cell clusters embedded in the marrow, was found in 46 cases (43.4%). The infiltrating depth varied from 1 to 4 mm (mean, 2.6 ± 0.7 mm). No tumor cells were observed in the normal bone marrow areas next to the interface. CONCLUSIONS:: The transition from osteosarcoma tissue to bone marrow after neoadjuvant chemotherapy can be divided into two histological patterns: clear and infiltrated. The greatest infiltration width was 4 mm from tumor to normal marrow in this study. This depth should be considered in the presurgical plan.

The Surgical Treatment and Outcome of Nonmetastatic Extremity Osteosarcoma with Pathological Fractures.

BACKGROUND: Recent studies have suggested that the presence of a pathological fracture does not impact on oncologic outcomes and the feasibility of limb salvage surgery (LSS) in appropriately selected patients when combined with neoadjuvant chemotherapy. These have largely been single institutional studies with limited numbers. The Eastern Asian Musculoskeletal Oncology Group reviewed the data from three large volume Asian orthopedic oncology centers to determine whether the presence of a pathologic fracture affected outcomes in osteosarcoma patients. METHODS: A retrospective review of the data was conducted. Ninety-five cases of nonmetastatic extremity osteosarcoma with a pathological fracture and 887 cases without fracture treated during the same period were compared. RESULTS: In the fracture group, the LSS rate was 62.1%, and the rate of amputation was 37.9%. In the nonfracture group, the LSS rate was 74.7%, and the amputation was 25.3%. In patients with a pathologic fracture, the rate of local recurrence for LSS and amputation groups was 8.5% and 2.8%, respectively. In this group, the 5-year survival in the LSS group was 66% as against. 46.8% in the amputation group. CONCLUSIONS: Our study suggests that surgically treated patients with pathologic fractures in osteosarcoma have adequate local control and do not have a poorer outcome compared to patients without a fracture. Though osteosarcoma with a pathologic fracture is not a contraindication for limb salvage, appropriate case selection is important when deciding local control options to ensure adequate oncologic clearance.

Radiofrequency ablation under 3D intraoperative Iso-C C-arm navigation for the treatment of osteoid osteomas.

OBJECTIVE: To evaluate the efficacy of intraoperative three-dimensional (3D) Iso-C C-arm-navigated percutaneous radiofrequency ablation (RFA) of osteoid osteomas. METHODS: 35 patients (20 males and 15 females) with osteoid osteomas underwent treatment with intraoperative 3D Iso-C C-arm navigation-guided RFA. The tumour was first biopsied for pathological examination, the core needle was removed and the RFA needle was inserted into the nidus. Post-operative X-rays and CT scans were performed to evaluate the degree of ablation and to assess for recurrence at 3-month follow-up. Patients also completed a visual analogue scale (VAS) both pre-operatively and 3 days post-operatively to subjectively assess pain. RESULTS: Pathological diagnosis confirmed osteoid osteoma in 19 cases. The other 16 cases were not pathologically diagnosed owing to inadequate biopsy specimens. In all cases, localized pain was immediately relieved following RFA. Patients reported significantly decreased pain, with mean pre-operative VAS scores of 3.4 reducing to 0.80 at 3 days post-operatively and further to 0.06 at 3-month follow-up (p < 0.05). The mean follow-up time was 15.5 months (range: 3-38 months). CONCLUSION: 3D Iso-C C-arm navigation-guided RFA is a safe and effective option for the treatment of osteoid osteomas and may be considered in place of intraoperative CT-guided and open resection. ADVANCES IN KNOWLEDGE: C-arm image-guided percutaneous RFA mitigates the need for pre-operative CT as well as intraoperative scintigraphy, provides real-time imaging of the anatomy, facilitates accurate resection of the tumour and enables immediate confirmation of excision.

Proteomics analysis reveals post-translational mechanisms for cold-induced metabolic changes in Arabidopsis.

Cold-induced changes of gene expression and metabolism are critical for plants to survive freezing. Largely by changing gene expression, exposure to a period of non-freezing low temperatures increases plant tolerance to freezing-a phenomenon known as cold acclimation. Cold also induces rapid metabolic changes, which provide instant protection before temperature drops below freezing point. The molecular mechanisms for such rapid metabolic responses to cold remain largely unknown. Here, we use two-dimensional difference gel electrophoresis (2-D DIGE) analysis of sub-cellular fractions of Arabidopsis thaliana proteome coupled with spot identification by tandem mass spectrometry to identify early cold-responsive proteins in Arabidopsis. These proteins include four enzymes involved in starch degradation, three HSP100 proteins, several proteins in the tricarboxylic acid cycle, and sucrose metabolism. Upon cold treatment, the Disproportionating Enzyme 2 (DPE2), a cytosolic transglucosidase metabolizing maltose to glucose, increased rapidly in the centrifugation pellet fraction and decreased in the soluble fraction. Consistent with cold-induced inactivation of DPE2 enzymatic activity, the dpe2 mutant showed increased freezing tolerance without affecting the C-repeat binding transcription factor (CBF) transcriptional pathway. These results support a model that cold-induced inactivation of DPE2 leads to rapid accumulation of maltose, which is a cold-induced compatible solute that protects cells from freezing damage. This study provides evidence for a key role of rapid post-translational regulation of carbohydrate metabolic enzymes in plant protection against sudden temperature drop.

[Surgical treatment of massive soft tissue sarcoma in the shoulder girdle].

OBJECTIVE: To detect the character of surgical treatment of massive soft tissue sarcoma in the shoulder girdle and analyze the impact factor to the result. METHODS: Seven patients with massive soft tissue sarcoma in the shoulder girdle were treated in our department between 2005 and 2009. There were 4 males and 3 females. All the patients were referred to our hospital after local recurrence post-operatively. The mean age was 43.8 years old (range 14 - 75). The maximum diameter of the tumor varied from 10 to 16 centimeters. All the patients were performed surgery, wide margin in 4 cases and marginal margin in 3 cases. Five were performed tumor resection and reconstruction with latissimus dorsi muscle flap transfer and skin graft. One was reconstructed with advanced skin flap and skin graft. The other one was treated with skin graft. The diagnosis included 3 malignant fibrous histiocytomas, 1 low grade myxoid fibrosarcoma, 1 Primitive neuroectodermal tumor, 1 rhabdomyosarcoma, 1 dermatofibrosarcomas protuberans. The MSTS score system was used to evaluate the shoulder function. RESULTS: Seven patients were followed up with long time. The mean follow up was 29 months (range 10 to 46 months). Two patients suffered local recurrence and one died of pulmonary metastasis 6 months after the second surgery for local recurrence. One patient suffered pulmonary metastasis. The last four patients were disease-free at the end of follow-up. The function of shoulder girdle was satisfactory. The mean MSTS score was 28. CONCLUSIONS: Soft tissue sarcomas in the shoulder girdle are easy to be misdiagnosed and mistreated. Wide surgical margin was the key impact factor to the local recurrence of soft tissue sarcoma in the shoulder girdle. The surgical margin and invasion of the tumor are the key factor to the prognosis. The soft tissue defect after surgery is often reconstructed by muscle flap transfer or skin flap transfer. The latissimus dorsi muscle flap transfer is often used.

Decreased expression of human aquaporin-5 correlated with mucus overproduction in airways of chronic obstructive pulmonary disease.

AIM: To investigate the relationship between aquaporin-5 (AQP5) expression and mucus overproduction in the airways of Chinese patients with chronic obstructive pulmonary disease (COPD) and the correlation with pulmonary function change. METHODS: Bronchial tissues were obtained from fiberoptic bronchoscopy and bronchial biopsy in West China Hospital from April to July 2004. Twenty-five patients were diagnosed as COPD patients, and another 20 were diagnosed as the control patients. The expressions of AQP5, mucin 5AC (MUC5AC), and mucin in bronchial tissues were detected by semiquantitative RT-PCR, in situ hybridization, and immunohistochemical and alcian blue-periodic acid-Schiff (AB-PAS) staining, respectively. RESULTS: Compared with the control group, an attenuated expression of AQP5 was detected throughout the bronchial tissues from patients with COPD (P<0.01), but no difference existed in the lung tissues (P>0.05). Simultaneously, increased staining of MUC5AC and mucus in submucosal glands were noted (P<0.01, respectively). Smoking attenuated the expressions of AQP5 and increased the staining of MUC5AC and mucus in submucosal glands in the COPD groups (P<0.01), while there were no significant differences observed in the control group (P>0.05). The decreased expression of AQP5 mRNA was correlated with forced expiratory volume in 1 s (FEV1)/forced vital capacity (FVC) (r=0.60, P<0.01), FEV1% predicted value (r=0.60, P<0.01), maximal expiratory flow in 50% vital capacity (V50) % predicted value (r=0.55, P<0.01), and maximal expiratory flow in 25% vital capacity (V25) % predicted value (r=0.45, P<0.01). The decreased expression was negatively correlated with MUC5AC mRNA of the epithelium airways (r=-0.45, P<0.01) and the AB-PAS-stained area of submucosal glands (r=-0.61, P<0.01). The upregulation of MUC5AC mRNA correlated with the positively AB-PAS-stained area of submucosal glands and correlated negatively with FEV1/FVC (r=-0.53; P<0.01), FEV1% predicted value (r=-0.53; P<0.01), V50% predicted value (r=-0.48; P<0.01), and V25% predicted value (r=-0.43; P<0.01). CONCLUSION: The attenuated gene expression of AQP5 existed in the airways of Chinese COPD patients, which was complicated by mucus hypersecretion. The decreased expression of AQP5 mRNA may be related to the severity of airflow obstruction.

Analysis of variabilities of serum proteomic spectra in patients with gastric cancer before and after operation.

AIM: To study the variabilities of serum proteomic spectra in patients with gastric cancer before and after operation in order to detect the specific protein markers that can be used for quick diagnosis of gastric cancer. METHODS: Proteomic spectra of 46 serum samples from patients with gastric cancer before and after operation and 40 from normal individuals were generated by IMAC-Cu protein chip and surface-enhanced laser desorption/ ionization time of flight mass spectrometry. RESULTS: Fourteen differentially expressed proteins in serum were screened by analysis of proteomic spectra of preoperative patients and normal individuals. We obtained 4 proteins (heat shock protein 27, glucose-regulated protein, prohibitin, protein disulfide isomerase A3) making up marker pattern which was able to class the patient-team and normal-team. These marker patterns yielded 95.7% sensitivity and 92.5% specificity, respectively. The proteins over-expressed in serum of preoperative patients were obviously down-regulated. CONCLUSION: Specific protein markers of gastric cancer can be used for the quick diagnosis of gastric cancer and judgment of prognosis. SELDI-TOF-MS is a useful tool for the detection and identification of new protein markers in serum.