Clinical laboratory characteristics and gene mutation spectrum of Ph-negative MPN patients with atypical variants of JAK2, MPL, or CALR.

OBJECTIVE: To evaluate the incidence, clinical laboratory characteristics, and gene mutation spectrum of Ph-negative MPN patients with atypical variants of JAK2, MPL, or CALR. METHODS: We collected a total of 359 Ph-negative MPN patients with classical mutations in driver genes JAK2, MPL, or CALR, and divided them into two groups based on whether they had additional atypical variants of driver genes JAK2, MPL, or CALR: 304 patients without atypical variants of driver genes and 55 patients with atypical variants of driver genes. We analyzed the relevant characteristics of these patients. RESULTS: This study included 359 patients with Ph-negative MPNs with JAK2, MPL, or CALR classical mutations and found that 55 (15%) patients had atypical variants of JAK2, MPL, or CALR. Among them, 28 cases (51%) were male, and 27 (49%) were female, with a median age of 64 years (range, 21-83). The age of ET patients with atypical variants was higher than that of ET patients without atypical variants [70 (28-80) vs. 61 (19-82), p = 0.03]. The incidence of classical MPL mutations in ET patients with atypical variants was higher than in ET patients without atypical variants [13.3% (2/15) vs. 0% (0/95), p = 0.02]. The number of gene mutations in patients with atypical variants of driver genes PV, ET, and Overt-PMF is more than in patients without atypical variants of PV, ET, and Overt-PMF [PV: 3 (2-6) vs. 2 (1-7), p < 0.001; ET: 4 (2-8) vs. 2 (1-7), p < 0.05; Overt-PMF: 5 (2-9) vs. 3 (1-8), p < 0.001]. The incidence of SH2B3 and ASXL1 mutations were higher in MPN patients with atypical variants than in those without atypical variants (SH2B3: 16% vs. 6%, p < 0.01; ASXL1: 24% vs. 13%, p < 0.05). CONCLUSION: These data indicate that classical mutations of JAK2, MPL, and CALR may not be completely mutually exclusive with atypical variants of JAK2, MPL, and CALR. In this study, 30 different atypical variants of JAK2, MPL, and CALR were identified, JAK2 G127D being the most common (42%, 23/55). Interestingly, JAK2 G127D only co-occurred with JAK2V617F mutation. The incidence of atypical variants of JAK2 in Ph-negative MPNs was much higher than that of the atypical variants of MPL and CALR. The significance of these atypical variants will be further studied in the future.

DNA-Compatible Cyclization Reaction to Access 1,3,4-Oxadiazoles and 1,2,4-Triazoles.

DNA-encoded chemical library (DECL) technology is a commonly employed screening platform in both the pharmaceutical industry and academia. To expand the chemical space of DECLs, new and robust DNA-compatible reactions are sought after. In particular, DNA-compatible cyclization reactions are highly valued, as these reactions tend to be atom economical and thus may provide lead- and drug-like molecules. Herein, we report two new methodologies employing DNA-conjugated thiosemicarbazides as a common precursor, yielding highly substituted 1,3,4-oxadiazoles and 1,2,4-triazoles. These two novel DNA-compatible reactions feature a high conversion efficiency and broad substrate scope under mild conditions that do not observably degrade DNA.

Expression analysis of TRAF2­ and NCK­interacting protein kinase (TNIK) and phosphorylated TNIK in papillary thyroid carcinoma.

The aim of the present study was to evaluate the expression of TRAF2- and NCK-interacting kinase (TNIK) and the levels of the active form of TNIK, phosphorylated (p)-TNIK, in papillary thyroid carcinoma (PTC), and to identify and compare the levels of TNIK and p-TNIK among PTC, benign thyroid tumors and normal tissues. The levels of TNIK and p-TNIK were examined by reverse transcription-quantitative (RT-q)PCR and immunohistochemical analysis (IHC) in PTC, benign thyroid tumors and normal tissues, and their association with clinicopathological features was evaluated. First, analysis of the Gene Expression Profiling Interactive Analysis and The Cancer Genome Atlas datasets suggested that the mRNA expression of TNIK was markedly increased in PTC tissues compared with that in normal tissues. RT-qPCR analyses then indicated that the relative mRNA expression of TNIK in PTC tissues was 4.47±6.16, which was significantly higher than that in adjacent tissues 2.57±5.83. The IHC results suggested that the levels of TNIK and p-TNIK in PTC tissues were markedly elevated compared with those in benign thyroid tumors and normal tissues. The levels of p-TNIK in patients with PTC were significantly associated with extrathyroidal extension (χ2=4.199, P=0.040). Positive staining for TNIK was observed in 187 out of 202 (92.6%) cases in the cytoplasm, nucleus or cytomembrane of PTC cells. Among the 187 positive cases, cytoplasm expression was identified in 162 cases (86.6%), nuclear expression in 17 cases (9.1%) and cytomembrane expression in 8 cases (4.3%). Positive staining for p-TNIK was observed in 179 out of 202 (88.6%) cases in the nuclei, cytoplasm or cytomembrane of PTC cells. In the 179 p-TNIK-positive cases, localization in the nuclei plus cytoplasm was identified in 142 cases (79.3%), nuclear localization in 9 cases (5.0%), presence in the cytoplasm in 21 cases (11.7%) and cytomembrane localization in 7 cases (3.9%). Both TNIK and p-TNIK were upregulated in PTC tissues and p-TNIK was significantly associated with extrathyroidal extension. It may act as a crucial oncogene to participate in PTC carcinogenesis and progression.

circ_0052184 Promotes Colorectal Cancer Progression via Targeting miR-604/HOXA9 Axis.

Background: The mortality rate of colorectal cancer (CRC) ranks second. circRNAs are abnormal expression in some diseases, and their dysregulation is associated with cancer progression. Recent studies have shown that the malignant progression of colorectal cancer is inseparable from the abnormal expression of circRNAs. Methods: First, the circ_0052184 expression in clinical tissue and cell samples was analyzed by qRT-PCR. Then, we constructed circ_0052184-silenced CRC cells and detected by qRT-PCR. Furthermore, the proliferation ability of cells was detected by colony formation assay. Cell migration ability was tested by wound healing assay and transwell assay. Cell invasion ability was detected by transwell assay. Results: Expression of circ_0052184 was significantly increased in colorectal cancer cell lines and tissues. Silencing circ_0052184 affected the proliferation, migration, and invasion of colorectal cancer cells. miR-604 was targeted by circ_0052184. The downstream target of miR-604 was HOXA9, and silencing circ_0052184 inhibited HOXA9 expression. The existence of the circ_0052184/miR-604/HOXA9 regulatory network in colorectal cancer was validated. circ_0052184 promoted the occurrence and development of colorectal cancer by targeting the miR-604/HOXA9 axis. Conclusions: Our study revealed that the molecular mechanism of circ_0052184 regulated the miR-604/HOXA9 axis, which might promote the malignant progression of colorectal cancer cells.

Spontaneous Motion and Rotation of Acid Droplets on the Surface of a Liquid Metal.

Self-propulsion of droplets is of great significance in many fields. The spontaneous horizontal motion and self-jumping of droplets have been well realized in various ways. However, there is still a lack of an effective method to enable a droplet to rotate spontaneously and steadily. In this paper, by employing an acid droplet and a liquid metal, the spontaneous and steady rotation of droplets is achieved. For an acid droplet, it may spontaneously move when it is deposited on the surface of the liquid metal. By adjusting experimental parameters to the proper range, the self-rotation of droplet happens. This phenomenon originates from the fluctuation of the droplet boundary and the collective movement of bubbles that are generated by the chemical reactions between the acid droplet and liquid metal. This rotation has a simpler implementation method and more steady rotation state. Its angular velocity is much higher than that driven by other mechanisms. Moreover, the movements of acid droplets on the liquid metal are classified according to experimental conditions. The internal flow fields, the movements and distribution of bubbles, and the fluctuation of the droplet boundary are also explored and discussed. The theoretical model describing the rotational droplet is given. Our work may deepen the understanding of the physical system transition affected by chemical reactions and provide a new way for the design of potential applications, e.g., micro- and nanodevices.

A Reactive Oxygen Species (ROS) Activated Hydrogen Sulfide (H2S) Donor with Self-Reporting Fluorescence.

Hydrogen sulfide (H2S) is an important cellular signaling molecule, and its physiological and pathophysiological properties have been under intensive investigation. In this study, a novel ratiometric fluorescent H2S donor (HSD-B) has been developed, which exhibited the following advantages: (i) scavenging ROS and producing H2S simultaneously; (ii) providing ratiometric fluorescence for visualization and quantification of H2S releasing; and (iii) targeting mitochondrion specifically. Moreover, it demonstrated protective effects on myocardial ischemia reperfusion injury in a cellular model. These attractive features promise this HSD-B as a fluorescent H2S donor for future research studies.

The effect of sharp solid edges on the droplet wettability.

Understanding the role of pinning force in droplet dynamic wetting is of critical importance for surface science studies. Generally, the pinning force is only related to the surface tension and the change of contact angle. However, there is an obvious correlation between the pinning force and the surface geometry. In this paper, the relation between the surface geometry and contact line pinning was studied with systematical experiments and theoretical analysis. We compared the samples with different edge angles and carried out plenty experiments with different liquids. Meanwhile, the theoretical analysis and molecular simulation were carried out. The results show that the sharp edge has a strong pinning effect on the contact line and can significantly change the contact angle and wetting state of droplets. The maximum contact angle of droplet has a linear relation with the edge angle of substrate. The formula of pinning force was revised to consider the impact of surface topography. According to the relationship between surface defect and contact line, we proposed a model to classify the cases of contact line pinning for the first time. Our research will deepen the understanding of contact line pinning and provide help for potentially industrial production designs.

Polymeric micelles with dual thermal and reactive oxygen species (ROS)-responsiveness for inflammatory cancer cell delivery.

BACKGROUND: The object of this study was to develop a thermally and reactive oxygen species-responsive nanocarrier system for cancer therapy. RESULTS: PPS-PNIPAm block copolymer was designed and synthesised using a combination of living anionic ring-opening polymerization and atom transfer radical polymerization. The synthesized polymer formed micellar aggregates in water and demonstrated dual responsiveness towards temperature and oxidants. Using doxorubicin (DOX) as a model drug, encapsulation and in vitro release of the drug molecules in PPS-PNIPAm nanocarriers confirmed the responsive release properties of such system. Cell uptake of the DOX loaded micelles was investigated with human breast cancer cell line (MCF-7). The results showed Dox-loaded micelles were able to be taken by the cells and mainly reside in the cytoplasma. In the stimulated cells with an elevated level of ROS, more released DOX was observed around the nuclei. In the cytotoxicity experiments, the Dox-loaded micelles demonstrated comparable efficacy to free DOX at higher concentrations, especially on ROS stimulated cells. CONCLUSIONS: These results demonstrated that PPS-PNIPAm nanocarriers possess the capability to respond two typical stimuli in inflammatory cells: temperature and oxidants and can be used in anticancer drug delivery.

Total Syntheses of Anti-HIV Cyclodepsipeptides Aetheramides A and B.

A concise total synthesis of aetheramide A in an overall yield of 4.7% with a longest linear sequence of 15 steps is described. This synthetic strategy features macrocyclization via an intramolecular trapping of acylketene generated from dioxinone precursor, and stereoselective late-stage methylation of ß-ketoamide. Aetheramide B could be synthesized via the ester migration of aetheramide A.

Asymmetric Total Syntheses of Aetheramides and Their Stereoisomers: Stereochemical Assignment of Aetheramides.

The concise total syntheses of the potent HIV inhibitors aetheramides A and B (IC50 values of 15 and 18 nM), as well as three pairs of their stereoisomers, were achieved, which allowed the complete stereochemical assignment of aetheramides for the first time. With a longest linear sequence of 15 steps, the convergent, fully stereocontrolled route provided aetheramides A and B in 5.3% and 3.6% yields, respectively. The synthetic strategy features efficient Stille coupling for macrocyclization, asymmetric aldol reactions to establish the ambiguous stereochemistries at C-17 and C-26, and implementation of mild conditions to avoid the epimerization of the sensitive polyketide moiety and the migration of the labile lactone.

[Role of carbon nanoparticles in patients with thyroid carcinoma undergoing total thyroidectomy plus bilateral central neck dissection].

OBJECTIVE: To evaluate the role of carbon nanoparticles for dissecting lymph nodes and preserving parathyroid glands in patients with thyroid carcinoma undergoing total thyroidectomy plus bilateral central neck dissection. METHODS: A total of 86 patients with thyroid carcinoma undergoing primarily total thyroidectomy plus bilateral central neck dissection were randomly divided into trial and control groups. Carbon nanoparticles were injected into thyroid gland of trial group. Total lymph node, metastasis lymph node, black stained lymph node and black stained metastasis lymph node of trial group were counted in central compartment dissection specimens. And total lymph node and metastasis lymph node of control group were counted in central compartment dissection specimens. Parathyroid glands in central neck dissection specimens were counted in two groups. For two groups, serum total calcium and parathyroid hormone were measured pre-operation and 1 day, 3 days, 5 days, 1 month, 3 months and 6 months post-operation. RESULTS: In trial group, the average counts of (10.19 ± 4.27) lymph nodes and (8.44 ± 4.31) black stained lymph nodes were more than those in control group at (6.26 ± 2.98) lymph nodes (all P < 0.01). Parathyroid gland was found in trial group (n = 7) and control group (n = 11). And the difference had no statistical significance (χ(2) = 1.124, P = 0.289). The preoperative serum levels of calcium and parathyroid hormone decreased within 6 months postoperatively in both groups (all P < 0.01). Without extracapsular invasion, serum levels of calcium and parathyroid hormone were higher in trial group than those in control group at 5 days, 1 month and 3 months post-operation (all P < 0.05). In an event of lymph node metastasis, serum levels of calcium (t = 3.446, P = 0.001) and parathyroid hormone (t = 2.441, P = 0.017) in trial group were higher than those in control group at 1 month post-operation. When there was extracapsular invasion or no lymph node metastasis, the serum levels of calcium and parathyroid hormone had no inter-group statistical differences within 6 months post-operation (all P > 0.05). When tumor size was less than or equal to 4 cm, the level of parathyroid hormone was higher in trial group than that in control group at 1 month post-operation (t = 2.703, P = 0.009). But no inter-group statistical differences existed within 6 months post-operation (all P > 0.05) when tumor size surpassed 4 cm. Regardless of tumor size, the serum levels of calcium in trial group were higher than those in control group at 1 month post-operation (all P < 0.01). CONCLUSIONS: For thyroid carcinoma patients, total thyroidectomy plus bilateral central neck dissection increases the incidence of hypoparathyroidism. Lymph nodes of central compartment may be tagged by carbon nanoparticles so as to boost the detection rate of lymph node. In events of lymph node metastasis or no extracapsular invasion, carbon nanoparticles can adequately protect parathyroid functions.

[The role of medical imaging plus carbon nanoparticles to manage the cervical lymph nodes in patients with thyroid carcinoma].

OBJECTIVE: The purpose of this study was to discuss the role of the combination of carbon nanoparticles and medical imaging to manage the cervical lymph nodes in patients with thyroid carcinoma. METHOD: Eighty one patients with thyroid carcinoma that primary treated were divided into two groups: trial group and control group. Carbon nanoparticles were injected into the thyroid gland of trial group patients. Central compartment (level VI) dissection, levels IIl and IV dissection, lateral node (levels II-V) dissection were performed respectively in all the patients on the basis of medical imaging and pathology. Total lymph nodes, metastasis lymph nodes, black stained lymph nodes and black stained metastasis lymph nodes of trial group were counted respectively in different dissection specimens. Total lymph nodes and metastasis lymph nodes of control group were counted respectively in different dissection specimens. Parathyroid glands of thyroid or central compartment dissection specimens were counted in two groups. RESULT: In trial group, rate of staining lymph node was 80.0% in central neck dissection tissue, 54.9% in levels III and IV dissection specimen, 39.1% in lateral node dissection specimen. In central compartment dissection tissue, lymph nodes on average in control group were less than in trial group (3.03 ± 2.07 vs. 4.72 ± 2.97) (P < 0.01). The same was in levels III and lV dissection specimen (5.53 ± 3.78 vs. 10.29 ± 3.36) (P < 0.01). As for lateral node dissection specimen,there was no statistic difference in the two group (13.4 ± 9.67 vs. 14.56 ± 6.28) (P > 0.05). There was no statistic difference between control group and trial group for the metastasis lymph nodes in difference dissection specimens. Parathyroid gland was found in 3 thyroid or central compartment dissection specimens among trial group, which was found in 9 specimens among control group, the difference had statistical significance (P < 0.05). CONCLUSION: During levels III and IV dissection in cN0 patients or central compartment dissection, lymph nodes can be signed well by carbon nanoparticles, which can improve the lymph node detection rate, but can not increase the lymph node detection rate in cN+ patients. Parathyroid gland can be preserved by carbon nanoparticles during the thyroid gland resection and central neck dissection.

[The combination of ultrasound and CT evaluate lymph node metastasis of thyroid papillary carcinoma in different compartments].

OBJECTIVE: To evaluate the role of the combination of ultrasound and enhanced CT in analyzing lymph node metastasis in thyroid papillary carcinoma (PTC) patients by compartment. METHOD: Clinical data of 115 cases (141 sides) with PTC were collected. All had undergone ultrasound in neck and enhanced CT both in neck and in mediastinum before surgery. They were divided into ultrasound group. CT group, and the combination of ultrasound and enhanced CT group to evaluate lymph node metastasis. RESULT: For the central compartment, the accuracy of ultrasound was 61.0%. CT was 48.9%, and the combination of ultrasound and CT was 62.4%. For the lateral compartment, ultrasound was 87.9%, CT was 78.7%, the combination of ultrasound and CT was 85.8%. Ultrasound had higher accuracy than CT in the central (P < 0.05) and lateral (P < 0.05) compartment. The combination of ultrasound and CT had higher accuracy than CT in the central compartment (P < 0.05), but there was no significant difference in the lateral compartment (P > 0.05). There was no significant difference in accuracy between ultrasound and the combination of ultrasound and CT neither in central (P > 0.05) nor in lateral (P > 0.05) compartment. Six cases of lymph node metastasis in mediastinum and 1 case in parapharyngeal space detected by CT were pathologically proven. CT found that five patients with pulmonary metastasis. CONCLUSION: The combination of ultrasound and CT or single ultrasound has higher accuracy in preoperative evaluation than single CT for lymph node metastasis in PTC. CT can assess some compartments such as mediastinum which can't be detected by ultrasound, and at the same time to evaluate lung metastasis. To evaluate lymph node metastasis in PTC, the combination of ultrasound and CT is more accurate and considerate than single method.

[Discussion about cervical incision of goiter in mediastinum posterior and its indications].

OBJECTIVE: To explore the feasibility of cervical approach for goiter in posterior mediastinum. METHOD: According to the mechanism that goiter filed into posterior mediastinum and the dissection of thyroid gland and mediastinum, we designed the following surgery principles (1) From top to bottom. (2) Find out recurrent nerve at the place where it enters larynx, then dissect recurrent nerve as long as possible and protect it carefully, meanwhile, search thyroid vessels along recurrent nerve. (3) To avoid and uncontrollable serious hemorrhage in the operation, all normal and aberrant blood vessels must be ligated cautiously, and avoid pulling great vessels in the thoracic part. (4) Separating tumor of hemorrhage under surgical capsule. Bluntly, it can avoid damage important structure in most occasions. (5) If the tumor of hemorrhage was difficult to be separated from the surrounding structure, ask thoracic surgeon for cooperation. RESULT: Two operations case were operated under the above guideline successfully, and the operations were performed with satisfactory effect, minimal invasion, rapid recovery and low medical cost. CONCLUSION: Cervical approach for goiter in mediastinum posterior is an ideal method of surgery, but it has following operative indication. (1) imaging date indicate that tumor of posterior septum is not connected to the surrounding structure. (2) It is not accompanied with superior vena cava syndrome. (3) The size of large thyroid tumor of posterior septum could be decreased by taking out the center part of tumor, and it is suitable for liquidized center tissue especially, then take out the tumor from neck. If it is hard to be taken out, you can ask thoracic surgeon for help.

[Phonation reconstruction and function assessment after total and near-total laryngectomy].

BACKGROUND & OBJECTIVE: Total laryngectomy is a common surgery pattern for treating laryngeal and hypopharyngeal cancer, but patients will lost phonation function after operation. This study was to retrospectively analyze usage of modified Amatsu, and modified Pearson operation in treating laryngeal and hypopharyngeal cancers, and to analyze phonation function by subjective and objective assessment. METHODS: Clinical data of 69 patients with laryngeal or hypopharyngeal cancer treated by total or near-total laryngectomy (modified Amatsu or modified Pearson operation) in our department from 1996 to 2003 were retrospectively analyzed. Phonation function of 69 patients was evaluated by acoustic analysis and Jiyan classification method, and compared with phonation quality of patients received vertical hemilaryngectomy (VHL) and healthy people. RESULTS: No patient appeared obvious aspiration. Four cases with post-operative radiotherapy all regained phonation function. The 3-year survival rate was 88% (22/25); 5-year survival rate was 80% (4/5). Results of acoustic assessment, and Jiyan classification method indicated that phonation function of patients received modified Amatsu or modified Pearson operation could meet daily requirements. CONCLUSIONS: Modified Amatsu and modified Pearson operations can be learnt and applied easily for phonation reconstruction after laryngectomy. They may be optional surgery patterns, especially modified Pearson operation.