Differential Diagnosis of Pathological Type of Peripheral Lung Cancer with Multimodal Contrast-Enhanced Ultrasound.

OBJECTIVE: The goal of the work described here was to investigate the role of multimodal contrast-enhanced ultrasound in the differential diagnosis of peripheral lung cancer. METHODS: From April 2017 to July 2021, 109 patients with confirmed pulmonary malignant lesions who underwent CEUS examination were involved in our study. Seven patients were excluded because of the short duration of CEUS video or unsatisfactory imaging. Finally,102 patients with peripheral lung cancer were enrolled in this study. The maximum diameter of the lesions ranged from 1.6 to 13.0 cm (mean 6.2 ± 2.3 cm). On the basis of the pathological results, the patients were divided into the small cell lung cancer (SCLC) group and non-small cell lung cancer (NSCLC) group (including adenocarcinoma, lung squamous cell carcinoma and large cell neuroendocrine carcinoma). A Logiq E9 ultrasonic machine equipped with a 3.5 to 5.0 MHz C5-1 probe was used. Patient clinical information, CEUS features, CPI patterns and TIC parameters were analyzed and compared between different groups. Statistical analyses were performed with SPSS software and MedCalc software. The receiver operating characteristic curve was plotted. RESULTS: In the differential diagnosis of SCLC and NSCLC, color parametric imaging indicated great performance. NSCLC exhibited a centripetal enhancement pattern more frequently (72.7%), while SCLC exhibited an eccentric enhancement pattern more frequently (92.9%) (p < 0.001). In the differential diagnosis of adenocarcinoma and squamous cell carcinoma, logistic regression analysis revealed that patient age of onset ≤60 y, difference in arrival time between lung and tumor ≤3.8 s, drop time of the time-intensity curve >23.2 s and absence of internal necrosis on CEUS were independent predictors for adenocarcinoma (area under the curve = 0.861). CONCLUSION: In our study, multimodal contrast-enhanced ultrasound provided useful information in the differential diagnosis between small cell lung cancer and non-small cell lung cancer, especially between adenocarcinoma and squamous cell carcinoma.

Fangchinoline inhibits mouse oocyte meiosis by disturbing MPF activity.

Fangchinoline (FA) is an alkaloid derived from the traditional Chinese medicine Fangji. Numerous studies have shown that FA has a toxic effect on various cancer cells, but little is known about its toxic effects on germ cells, especially oocytes. In this study, we investigated the effects of FA on mouse oocyte maturation and its potential mechanisms. Our results showed that FA did not affect meiosis resumption but inhibited the first polar body extrusion. This inhibition is not due to abnormalities at the organelle level, such as chromosomes and mitochondrial, which was proved by detection of DNA damage and reactive oxygen species. Further studies revealed that FA arrested the oocyte at the metaphase I stage, and this arrest was not caused by abnormal kinetochore-microtubule attachment or spindle assembly checkpoint activation. Instead, FA inhibits the activity of anaphase-promoting complexes (APC/C), as evidenced by the inhibition of CCNB1 degeneration. The decreased activity of APC/C may be due to a reduction in CDC25B activity as indicated by the high phosphorylation level of CDC25B (Ser323). This may further enhance Maturation-Promoting Factor (MPF) activity, which plays a critical role in meiosis. In conclusion, our study suggests that the metaphase I arrest caused by FA may be due to abnormalities in MPF and APC/C activity.

Metastasis of lung cancer?

BACKGROUND: Sarcoidosis is a multi-system, idiopathic, inflammatory disorder that affects the lungs in over 90% of patients. The incidence of bone lesions in sarcoidosis is only 1-13%. CASE REPORT: This study describes a 60-year-old woman with a previous history of thyroid cancer, and a more recent diagnosis of lung cancer with suspicious metastatic lesions, which were confirmed to be sarcoidosis. CONCLUSION: This case suggests that pulmonary neoplasms and pulmonary sarcoidosis can coexist and be easily confused. When lung cancer is accompanied by symmetric hilar lymph node enlargement and multiple lung nodules, sarcoidosis should be considered in addition to metastasis, and a biopsy should be performed for confirmation.

Axial and frontal X-ray fluoroscopy technique of the sustentaculum tali can improve the accuracy of sustentacular screw placement.

INTRODUCTION: Calcaneal fractures, especially those involving the articular surface, should be anatomically reduced as much as possible. Fixing the fracture by placing a screw into the sustentaculum tali from the lateral side of the calcaneus is generally considered to be the key to successful surgery. However, due to the limited visibility during surgery, it is not easy to place screws into the sustentaculum tali accurately. The purpose of this study was to explore a new fluoroscopy method for the sustentaculum tali and verify the value of this method in improving screw placement accuracy. METHODS: In this study, a total of 42 human foot and ankle specimens were dissected and measured. The shape and position of the sustentaculum tali were observed, and the influence of adjacent bones on imaging findings was analysed. The axial and frontal X-ray fluoroscopy method to view the sustentaculum tali was formulated, and the appropriate projection angle through anatomical and image measurements was explored. Thirty specimens were randomly selected for screw placement, and the direction of the screw was dynamically adjusted under the new imaging method. The success rate of sustentacular screw placement was evaluated. RESULTS: The anteversion angles of the sustentaculum tali were 30.81 ± 2.21° and 30.68 ± 2.86° by anatomical and imaging measurements, respectively. There was no statistically significant difference in the anteversion angle between the two measurement methods. Harris heel views should be obtained at 30° to identify the sustentaculum tali on axial X-ray images. Frontal X-ray imaging was performed perpendicular to this projection angle. Through frontal and axial X-ray imaging, the position and shape of the sustentaculum tali can be clearly observed, and these factors are seldom affected by adjacent bones. Under the new fluoroscopy method, the screws were placed from the anterior region of the lateral wall of the calcaneus to the sustentaculum tali. A total of 60 screws were placed in the 30 specimens; of these, 54 screws were in good position, 2 screws penetrated the cortical bone, and 4 screws did not enter the sustentaculum tali. The success rate of sustentacular screw placement was 90% (54/60). CONCLUSIONS: Axial and frontal X-ray images of the sustentaculum tali can clearly show the shape of the structure, which improves sustentacular screw placement accuracy.

CD4+ CD25+ regulatory T cells promote hepatocellular carcinoma invasion via TGF-ß1-induced epithelial-mesenchymal transition.

BACKGROUND: CD4+ CD25+ regulatory T cells (Tregs), a crucial component of the infiltration of immune cells in tumor microenvironment, are associated with progression and metastasis of hepatocellular carcinoma (HCC). METHODS: The mechanism of Tregs in the invasion and metastasis of HCC was investigated in vivo and in vitro using immunohistochemical analysis, western blot, and quantitative reverse transcription-PCR (qRT-PCR). RESULTS: Analysis of 78 clinical HCC samples indicated that high expression of Tregs was strongly associated with poor cancer-free survival and overall survival of patients. The reduced expression of E-cadherin and enhanced expression of Vimentin and transforming growth factor-beta 1 (TGF-ß1) were found in HCC tissue compared with normal liver tissue. The HCC Hepa1-6 cells were treated with the supernatant of Tregs-conditioned medium (Tregs-CM) to investigate the epithelial-mesenchymal transition (EMT) and TGF-ß1. Western blot and qRT-PCR also showed that down-regulated E-cadherin and up-regulated Vimentin and TGF-ß1 were found in Tregs-CM-treated Hepa1-6 cells. An experiment of tumorigenicity in C57 mice showed larger and heavier tumors in Tregs-CM-treated group than in the control group. Tregs produced higher TGF-ß1 compared with Tregs treated with FOXP3 shRNA. TGF-ß1 with neutralizing antibodies was used to deplete TGF-ß1 in Tregs-CM, which enhanced expression of E-cadherin, reduced expression of Vimentin and TGF-ß1, and decreased migratory and invasive capacity of Hepa1-6 cells. CONCLUSION: Tregs could promote the invasion and migration of Hepa1-6 cells, which are possibly maintained by TGF-ß1-induced EMT. This study showed that the development of therapeutic strategies against TGF-ß1 pathway is valuable in HCC therapy.

Pharmacokinetic and tissue distributions study of adenosine, 4-hydroxybenzyl alcohol and Parishin C from Gastrodia elata extract in rats.

The plant Gastrodia elata is a type of the orchid plant Gastrodia elata Bl. which contains glycosides, phenols, polysaccharides, sterols, and organic acids and a variety of active ingredients are proved to have certain pharmacological activities. To understand the process in the body of Gastridua elata, we used HPLC to study pharmacokinetics and tissue distributions of adenosine, 4-hydroxybenzyl alcohol and Parishin C in rats. The results showed that the three ingredients could be detected in plasma and different organizations at various time points. There was no significant difference in systemic clearance at three ingredients and it may be show that the three ingredients distributed (0.475±0.025, 0.518±0.033, 0.699±0.051) quickly and eliminated (5.37±0.87, 4.54±0.69, 5.34±0.82) slowly in plasma. There was the highest content of adenosine in spleen, followed by liver and lung. The highest content of 4-hydroxybenzylacohol in liver, and was higher in spleen. Parishin C was highest in heart, followed by liver and spleen. It is obvious that the contents of three ingredients are all higher in liver. The trends of the three ingredients' contents in G. rhizome extract were consistent with the contents in the plasma after intravenous administration.

Kinsenoside screening with a microfluidic chip attenuates gouty arthritis through inactivating NF-κB signaling in macrophages and protecting endothelial cells.

Gouty arthritis is a rheumatic disease that is characterized by the deposition of monosodium urate (MSU) in synovial joints cause by the increased serum hyperuricemia. This study used a three-dimensional (3D) flowing microfluidic chip to screen the effective candidate against MSU-stimulated human umbilical vein endothelial cell (HUVEC) damage, and found kinsenoside (Kin) to be the leading active component of Anoectochilus roxburghi, one of the Chinese medicinal plant widely used in the treatment of gouty arthritis clinically. Cell viability and apoptosis of HUVECs were evaluated, indicating that direct Kin stimulation and conditioned medium (CM) from Kin-treated macrophages both negatively modulated with MSU crystals. Additionally, Kin was capable of attenuating MSU-induced activation of nuclear factor-κB/mitogen-activated protein kinase (NF-κB/MAPK) signaling, targeting IκB kinase-α (IKKα) and IKKß kinases of macrophages and influencing the expressions of NF-κB downstream cytokines and subsequent HUVEC bioactivity. Inflammasome NLR pyrin domain-containing 3 (NALP3) and toll-like receptor 2 (TLR2) were also inhibited after Kin treatment. Also, Kin downregulated CD14-mediated MSU crystals uptake in macrophages. In vivo study with MSU-injected ankle joints further revealed the significant suppression of inflammatory infiltration and endothelia impairment coupled with alleviation of ankle swelling and nociceptive response via Kin treatments. Taken together, these data implicated that Kin was the most effective candidate from Anoectochilus roxburghi to treat gouty arthritis clinically.

[Thrombocytopenia induced by lipopolysaccharide may be not related to coagulation and inflammatory response].

OBJECTIVE: To explore the relationship between thrombocytopenia (TCP) induced by lipopolysaccharide (LPS) and coagulation or inflammatory response in mouse. METHODS: Forty-eight C57BL/6 mice were divided into control group, low-dose, and high-dose LPS treatment groups by random number table method, and each group was subdivided into 4-hour and 24-hour subgroups randomly, with 8 mice in each subgroup. 0.5 mg/kg or 50 mg/kg LPS was injected intraperitoneally in low-dose or high-does group respectively, and equal amount of normal saline was injected in control group. Blood was collected from endocanthal vein at the specified time point, platelet count (PLT) was counted, and the levels of thrombin antithrombin complex (TAT), D-dimer, fibrinogen degradation product (FDP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were determined by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with control group, PLT ( x 10(9)/L) at 4 hours and 24 hours in low-dose and high-dose LPS groups was significantly decreased (4 hours: 660.65 ± 180.48, 568.55 ± 117.99 vs. 1 199.13 ± 110.54; 24 hours: 505.63 ± 218.92, 256.33 ± 72.86 vs. 1 229.13 ± 1 189.37, all P < 0.05), and the changes were more obvious in high-dose LPS group compared with those of the low-dose LPS group (all P < 0.05). Factorial analysis showed that the changes in PLT were related with LPS dosage and time (F1 = 135.660, P = 0.000; F2 = 12.120, P2 = 0.001). It was also found that there was an interactive effect of the dose of LPS and time on PLT (F = 5.580, P = 0.007). Compared with control group, TAT, TNF-α, and IL-6 at 4 hours and 24 hours in low-dose and high-dose LPS groups were significantly decreased [TAT (ng/L) at 4 hours: 1.10 ± 0.59, 0.22 ± 0.13 vs. 3.47 ± 1.73; 24 hours: 1.18 ± 0.68, 0.39 ± 0.29 vs. 3.19 ± 1.27; TNF-α (nmol/L) at 4 hours: 87.35 ± 12.29, 93.70 ± 5.25 vs. 101.59 ± 10.96, 24 hours: 81.94 ± 8.26, 93.23 ± 4.71 vs. 102.84 ± 10.56; IL-6 (ng/L) at 4 hours: 81.78 ± 7.82, 78.59 ± 9.06 vs. 110.88 ± 9.66, 24 hours: 76.03 ± 9.85, 71.34 ± 3.69 vs. 110.88 ± 10.35, all P < 0.05]. TAT at 4 hours and 24 hours in high-dose LPS group was further decreased, and TNF-α at 24 hours was increased as compared with those of low-dose LPS group (all P < 0.05). TAT, TNF-α and IL-6 were influenced only by different dosage of LPS (TAT: F = 42.350, P = 0.000; TNF-α: F = 14.8 10, P = 0.000; IL-6: F = 81.910, P = 0.000), not time (TAT: F = 0.002, P = 0.967; TNF-α: F = 0.342, P = 0.562; IL-6: F = 2.973, P = 0.092). Changes in TAT was not found to be related with the dose of LPS and its time of action, or levels of TNF-α and IL-6 (TAT: F = 0.236, P = 0.791; TNF-α: F = 0.572, P = 0.569; IL-6: F = 0.774, P = 0.468). The dosage of LPS and time of admission showed no influence on D-dimer (F1 = 2.448, P = 0.099; F2 = 0.024; P2 = 0.877). The effect of different doses of LPS and time of administration showed no influence on FDP (F1 = 0.106, P = 0.900; F2 = 0.013, P2 = 0.908), and no interactive effects were found (D- dimer: F = 0.002, P = 0.998; FDP: F = 0.582, P = 0.563). CONCLUSION: LPS can induce TCP in mouse, but this effect may not related to the activation of coagulation system and excessive inflammatory response.

15d-PGJ2 inhibits cell growth and induces apoptosis of MCG-803 human gastric cancer cell line.

AIM: To investigate the influence of peroxisome proliferator-activated receptor gamma (PPARgamma) ligand, 15-deoxy-12, 14-prostaglandin J2 (15dPGJ2) on the proliferation and apoptosis of MCG-803 human gastric cancer cell lines. METHODS: Cell proliferation was measured by 3H-TdR assay. Apoptosis was determined by ELISA and TUNEL staining. Protein and mRNA level of bcl-2 family and COXs were measured by Western blotting and Northern blotting respectively. PGE2 production was examined by RIA. RESULTS: 15dPGJ2 inhibited cell growth and induced apoptosis of MCG-803 cells. The COX-2 and bcl-2/bax ratios were decreased following 15dPGJ2 treatment. The PGE2 production in supernatants was also decreased. These changes were in a dose-dependent manner. CONCLUSION: 15dPGJ2 may be a useful therapeutic agent for the treatment of gastric cancer.