Analysis of the resistance profile of real-world alectinib first-line therapy in patients with ALK rearrangement-positive advanced non-small cell lung cancer using organoid technology in one case of lung cancer.

Background: Alectinib has achieved excellent therapeutic efficacy in anaplastic lymphoma kinase (ALK) fusion gene-positive non-small cell lung cancer (NSCLC) patients, however, patients eventually develop resistance to it. Exploring the gene variant mapping after alectinib resistance provides a basis for the whole management of ALK-positive advanced NSCLC. This study aimed to characterize the mutation profiles of real-world ALK rearrangement-positive advanced NSCLC patients after first-line alectinib treatment resistance. The research also investigated the treatment options and coping strategies after resistance. Methods: Clinical data of patients with advanced NSCLC who received first-line alectinib treatment in the First Affiliated Hospital of Guangzhou Medical University between November 2018 and April 2022 were collected. Moreover, next-generation sequencing (NGS) data of the patient's baseline and post-resistance tissues were gathered. One patient underwent lung cancer organoid culture and drug sensitivity testing. Results: Out of 35 first-line alectinib-treated patients with advanced NSCLC, 31 are presently in progression-free survival (PFS; 4.3-35.0 months). Four patients experienced progressive disease, and all of them were sequentially treated with ceritinib. Tissue NGS results before sequential treatment in three patients indicated an echinoderm microtubule-associated protein-like 4-ALK fusion that remained at the original baseline, and the PFS for ceritinib treatment was 0.5-1.3 months. One patient developed acquired resistance mutations in the structural domain of ALK protein kinase (V1180L and E1161D), and the PFS for ceritinib treatment was 6.7 months. For one patient who maintained original baseline ALK rearrangement positive without acquired mutation after progression of ceritinib resistance, lung cancer-like organ culture with sequential brigatinib and lorlatinib led to a PFS of 3.2 and 1.9 months, respectively, which aligned with the corresponding drug susceptibility testing results for this patient. Conclusions: For ALK rearrangement-positive patients, blind sequencing of other second-generation tyrosine kinase inhibitors (TKIs) or third-generation lorlatinib may not guarantee satisfactory tumor suppression following first-line second-generation ALK-TKI alectinib administration for treatment progression. NGS testing of patients' blood or tissue samples after disease progression may provide insight into the etiology of alectinib resistance. Patient-sourced drug sensitivity testing of lung cancer-like organs selects drug-sensitive medications based on NGS results and provides a reference for subsequent drug therapy for patients after drug resistance, particularly those who remain ALK rearrangement-positive at baseline.

Chiral Discrimination of Acyclic Secondary Amines by 19F NMR.

Chiral aliphatic amine compounds exhibit a range of physiological activities, making them highly sought-after in the pharmaceutical industry and biological research. One notable obstacle in studying these compounds stems from the pronounced steric hindrance surrounding the nitrogen atom. This characteristic often leads to a weak affinity of acyclic secondary amines for molecular probes, making their chiral discrimination intricate. In response to this challenge, our research has unveiled a novel 19F-labeled probe adept at recognizing and distinguishing between enantiomers of these acyclic secondary amines. By strategically incorporating a single fluorine atom as the 19F label, we have managed to diminish the steric hindrance at the binding site. This alteration bolsters the probe's affinity toward bulkier analytes. As a testament to its effectiveness, we have successfully employed our probe in the chiral analysis of relevant pharmaceuticals, accurately determining their enantiocomposition.

Key immunity characteristics of diverse stages of brucellosis in rural population from Inner Mongolia, China.

BACKGROUND: Brucellosis poses a serious threat to human and animal health, particularly in developing countries such as China. The Inner Mongolia Autonomous Region is one of the most severely brucellosis-endemic provinces in China. Currently, the host immune responses functioning to control Brucella infection and development remain poorly understood. The aim of this study is to further clarify the key immunity characteristics of diverse stages of brucellosis in Inner Mongolia. METHODS: We collected a total of 733 blood samples from acute (n = 137), chronic (n = 316), inapparent (n = 35), recovery (n = 99), and healthy (n = 146) groups from the rural community of Inner Mongolia between 2014 and 2015. The proportions of CD4+, CD8+, Th1, Th2, and Th17 T cells in peripheral blood and the expression of TLR2 and TLR4 in lymphocytes, monocytes and granulocytes were examined using flow cytometry analysis. The differences among the five groups were compared using one-way ANOVA and the Kruskal-Wallis method, respectively. RESULTS: Our results revealed that the proportions of CD4+ and CD8+ T cells were significantly different among the acute, chronic, recovery, and healthy control groups (P < 0.05), with lower proportions of CD4+ T cells and a higher proportion of CD8+ T cells in the acute, chronic, and recovery groups. The proportion of Th1 cells in the acute, chronic, and inapparent groups was higher than that in the healthy and recovery groups; however, there was no significant difference between patients and healthy individuals (P > 0.05). The proportion of Th2 lymphocytes was significantly higher in the acute and healthy groups than in the inapparent group (P < 0.05). The proportion of Th17 cells in the acute group was significantly higher than that in the healthy control, chronic, and inapparent groups (P < 0.05). Finally, the highest expression of TLR4 in lymphocytes, monocytes and granulocytes was observed in the recovery group, and this was followed by the acute, chronic, healthy control, and inapparent groups. There was a significant difference between the recovery group and the other groups, except for the acute group (P < 0.05). Moreover, a correlation in TLR4 expression was observed in lymphocytes, monocytes and granulocytes among the five groups (r > 0.5), except for the inapparent group between lymphocytes and granulocytes (r = 0.34). CONCLUSIONS: Two key factors (CD8+ T cells and TLR4) in human immune profiles may closely correlate with the progression of brucellosis. The detailed function of TLR4 in the context of a greater number of cell types or tissues in human or animal brucellosis and in larger samples should be further explored in the future.

Assessment of Binding Interaction between Bovine Lactoferrin and Tetracycline Hydrochloride: Multi-Spectroscopic Analyses and Molecular Modeling.

In this paper, the interaction between bovine lactoferrin (bLf) and tetracycline hydrochloride (TCH) was researched by microscale thermophoresis (MST), multi-spectroscopic methods, and molecular docking techniques. Normal fluorescence results showed that TCH effectively quenched the intrinsic fluorescence of bLf via static quenching. Moreover, MST confirmed that the combination force between bLf and TCH was very strong. Thermodynamic parameters and molecular docking further revealed that electrostatic forces, van der Waals, and hydrogen bonding forces played vital roles in the interaction between bLf and TCH. The binding distance and energy transfer efficiency between TCH and bLf were 2.81 nm and 0.053, respectively. Moreover, the results of circular dichroism spectra (CD), ultraviolet visible (UV-vis) absorption spectra, fluorescence Excitation-Emission Matrix (EEM) spectra, and molecular docking verified bLf indeed combined with TCH, and caused the changes of conformation of bLf. The influence of TCH on the functional changes of the protein was studied through the analysis of the change of the bLf surface hydrophobicity and research of the binding forces between bLf and iron ion. These results indicated that change in the structure and function of bLf were due to the interaction between bLf and TCH.

Resveratrol inhibits age-dependent spontaneous tumorigenesis by SIRT1-mediated post-translational modulations in the annual fish Nothobranchius guentheri.

Resveratrol, SIRT1 activator, inhibits carcinogenesis predominantly performed in transgenic animal models, orthotopic cancers of nude mice or different cancer cell lines, but its effects during process of spontaneous tumors using vertebrate models remain untested. Spontaneous liver neoplasm is an age-related disease and is inhibited by resveratrol in the annual fish Nothobranchius guentheri, which indicates that the fish can act as an excellent model to study spontaneous tumorigenesis. Totally, 175 fish were fed with resveratrol and another 175 fish for controls. Treated fish were fed with resveratrol (25 µg/fish/day) from sexual maturity (4-month-old) until they were sacrificed at 6-, 9- and 12-month-old. Immunoblot, immunohistochemistry and co-immunoprecipitation were employed to investigate the underlying mechanisms that resveratrol inhibited age-dependent spontaneous tumorigenesis in the fish. Results showed that resveratrol increased protein level of SIRT1 and alleviated age-associated tumorigenesis in liver. With SIRT1 up-regulation, resveratrol reduced proliferation by deacetylating K-Ras and inactivating K-Ras/PI3K/AKT pathway; and promoted apoptosis through deacetylation and dephosphorylation of FoxOs, up-regulation of DLC1 and interaction between SIRT1 and DLC1, and dephosphorylation of DLC1 in spontaneous neoplasms. We established a novel short-lived fish model for understanding the molecular mechanisms of drugs on age-dependent spontaneous tumorigenesis.

Oogenesis, vitellogenin-mediated ovarian degeneration and immune response in the annual fish Nothobranchius guentheri.

Annual fishes of the genus Nothobranchius show expression of age-related biomarkers at behavioral and histological levels. They therefore represent an excellent animal model for aging studies. However, oocyte development, histological and biochemical degeneration and immune response of ovary in the annual fishes remain unclear. Here, using one of these short-lived fishes, Nothobranchius guentheri, we reported that oogenesis process was divided into four stages (oogonium, primary growth stage, cortical alveolus stage and vitellogenesis stage), and old ovaries showed histological degeneration (with decreased mature oocytes and increased atretic oocytes) accompaning with high levels of senescence-associated beta-galactosidase and lipofuscin by down-regulation of vitellogenin (the precursor of yolk proteins). Moreover, poly(I:C) induced inflammation with overexpression of NF-κB and IL-8, and up-regulated vitellogenin expression. It was a first analysis for vitellogenin to participate in ovarian degeneration and immune response in ovary of fish, indicating that vitellogenin fulfilled a critical role in ovary development and innate immune system.

Enhanced electrochemical performance of hydrous RuO2/mesoporous carbon nanocomposites via nitrogen doping.

Hydrous RuO2 nanoparticles have been uniformly deposited onto nitrogen-enriched mesoporous carbons (NMCs) via a facile hydrothermal method. The nitrogen doping in the carbon framework not only provides reversible pseudocapacitance but also guides uniform deposition of RuO2 nanoparticles. As a result, an extremely high specific capacitance of 1733 F/g per RuO2, comparable to the theoretic capacitance of RuO2, is reached when 4.3 wt % of RuO2·1.25H2O is loaded onto the NMCs. Systematic studies show that either nitrogen-free or excess nitrogen doping result in RuO2 clusters formation and worsen the electrochemical performances. With intermediate nitrogen and RuO2 content (8.1 wt % N, 29.6 wt % of RuO2·1.25H2O), the composites deliver excellent power performance and high specific capacitance (402 F/g) with reversible capacitive response at 500 mV/s. The unique properties of nitrogen in textual, morphological, and electrochemical aspects may also provide further understanding about the effects of nitrogen doping and metal oxide deposition on supercapacitor performance.

[Pathogenesis of liver fibrosis in patients with chronic hepatitis B].

OBJECTIVE: To explore the role of sinusoidal endothelial cell in the development of liver fibrosis, and to dissect the relationship among hepatic microcirculation disorders, hepatic sinusoidal capilarization and liver fibrosis. METHODS: Liver biopsy was performed in fifty-six patients with chronic hepatitis B. The liver tissues were observed under light microscope and transmitted electronic microscope. RESULTS: Of 56 cases, 39 cases were mild hepatitis, 10 were moderate hepatitis, and 7 were severe hepatitis. The morphology of hepatic stellate cells (HSCs) was similar to that of fibroblasts in the tissues of the patients with chronic hepatitis B. Collagenous fibers were deposited around the hepatic stellate cells. Electron-dense materials were deposited between sinusoidal endothelial cell and hepatic stellate cell. The size and amount of fenestraes of sinusoidal endothelial cells were reduced in 53 of 56 cases. The consecutive or inconsecutive membrane-like materials were observed along sinusoidal endothelial cells in 20 cases. Collagen fibers were observed in the space of Disse in 15 cases. Even in the patients with normal hepatic functions, red blood cells aggregation and microthrombi could be observed in the liver tissues. CONCLUSION: Sinusoidal endothelial cells are involved in development of liver fibrosis by interacting with hepatic stellate cells. Hepatic microcirculation disorders and sinusoidal capillarization are important changes in the early stage of liver fibrosis.

[Effects of flashing light on ocular growth and development of myopia in pigmented guinea pigs].

OBJECTIVE: To investigate the effects of flashing light exposure on ocular growth and development of myopia in guinea pigs. METHODS: Thirty 4-week-old pigmented guinea pigs were randomly divided into three groups. Animals in group I were reared with the flashing light continuously and lasted for 6 weeks. The flashing frequency was 15 times per minutes and every flash includes 2 seconds light on and 2 seconds dark. Animals in group II and III were reared with normal light on, and the illumination cycle was 12 hours light/12 hours dark in group II and 24 hours light in group III. After 6 weeks, the effects of flashing light on eye development were assessed by cycloplegic retinoscopy, a-scan ultrasonography and eye weight. The histopathology changes of sclera, choroids and retina in posterior pore of the eye were examined using light microscope and transmission electron microscope. RESULTS: At the end of the treatment period, guinea pigs reared in flashing light exhibited -7.00 D myopia, eye axial elongated 0.56 microm, and the weight increased 68 mg. The histopathology examination showed that the posterior sclera fibroblast become more active, the cell number increased, and the place between fibers became larger, the posterior choroids became thinned, and the outer membrane of photoreceptor cells became shorter and irregular, when compared with the control groups. CONCLUSION: Flashing light can promote ocular growth and induce myopia in pigmented guinea pigs.

Hepatic microcirculatory disturbances in patients with chronic hepatitis B.

OBJECTIVE: To document morphological changes in hepatic microcirculation in liver tissue with hepatitis B and the pathogenesis of hepatic microcirculatory disturbances. METHODS: Liver tissue samples were obtained from patients with hepatitis B by liver biopsy. These samples were examined with a light microscope and transmission electron microscope. RESULTS: Hepatic microcirculatory disturbances existed in patients with hepatitis B, including those with normal liver function, manifested by red blood cell aggregation in sinusoids seen under light microscope and sinusoidal capillarization seen under electron microscope. Weibel-Palade bodies in sinusoidal endothelial cells were seen in 26 out of 53 cases. Intimate contacts were found between lymphocyte/Kupffer cells and sinusoidal endothelial cells. CONCLUSIONS: Hepatic microcirculatory disturbances exist in patients with hepatitis B. The appearance of Weibel-Palade bodies in sinusoidal endothelial cells may be a key step in the development of hepatic microcirculatory disturbances.

[Role of placental apoptosis in fetal growth restriction].

OBJECTIVE: To determine the relationship of placental cellular apoptosis and pathophysiology of fetal growth restriction (FGR). METHODS: Placental samples were obtained from 18 pregnancies complicated by FGR and 14 normal pregnancies. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and transmission electron microscopy were used to confirm the occurrence of apoptosis. RESULTS: In FGR group the placental apoptosis rate was (n = 18) 12.1 per thousand, the average placental weight was (236 +/- 24) g, the average birth weight was (2,071 +/- 428) g; In normal group (n = 14), the placental apoptosis rate was 7.3 per thousand, the average placental weight was (354 +/- 63) g, the average birth weight was (3,411 +/- 588) g (P < 0.05). The incidence of apoptosis was significantly higher in placental samples from pregnancies with FGR compared with normal placental samples (P < 0.05). Under transmission election microscopy, apoptosis was obviously compact and the chromatins were formed as mass. CONCLUSION: These results suggest that apoptosis may play a role in the pathophysiologic mechanisms of FGR.