Y9, a Gboxin analog, displays anti-tumor effect in non-small cell lung cancer by inducing lysosomal dysfunction and apoptosis.

Non-small cell lung cancer (NSCLC) ranks among the most prevalent malignancies globally. Gboxin, a novel inhibitor of mitochondrial complex V that exerts unique anti-tumor effects via oxidative phosphorylation inhibition, but shows no efficacy against NSCLC in vivo. Through chemical structure optimization, we designed and synthesized Gboxin analog Y9, which demonstrates significantly enhanced potency over its predecessor. Specifically, Y9 inhibited NSCLC significantly more strongly than Gboxin and possessed the ability to inhibit cell cycle progression and induce oxidative stress similar to Gboxin. Further investigation revealed that unlike Gboxin, Y9 selectively acidifies lysosomes and induces lysosomal dysfunction. This leads to hyperactive autophagy with impaired substrate clearance, and ultimately resulting in apoptosis. Animal studies confirmed the efficacy of Y9 in suppressing tumor growth in a xenograft mouse model. Collectively, Y9 is a distinctive Gboxin analog that outperforms its prototype by inducing lysosomal dysfunction and apoptosis, and has the potential to be developed as a novel anti-NSCLC lead compound.

A novel Gboxin analog induces OXPHOS inhibition and mitochondrial dysfunction-mediated apoptosis in diffuse large B-cell lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is an aggressive B-cell non-Hodgkin's lymphoma. Currently, moderate efficacy and limitations of approved drugs still exist, and it is necessary to develop newer and more effective drugs. Gboxin is a promising inhibitor of OXPHOS, which specifically inhibits the growth of many kinds of cancer cell lines. In the present study, 21 Gboxin analogs incorporating amide and ester moieties were designed and synthesized. Preliminary screening results show that 5d also has specific selectivity for cancer cells, particularly on the DLBCL cells, which is weaker than that of Gboxin but still good. Thus, the effect and underlying mechanism of 5d on DLBCL cells were further studied. The results showed that 5d exhibits potent proliferation inhibition and cell cycle arrest effects, and its IC50 to DLBCL cells is below 1 µM. In addition, 5d induces apoptosis of DLBCL cells in a time- and dose-dependent manner, and this effect is stronger than that of Gboxin and VP16. Mechanistically, 5d plays its role mainly through the stimulation of metabolic stress in DLBCL cell lines, which induces OXPHOS inhibition, inflammation, DNA damage and mitochondrial dysfunction. These data suggest that 5d has potential as a candidate agent for DLBCL alternative drug development.

Chaetocochin J, an epipolythiodioxopiperazine alkaloid, induces apoptosis and autophagy in colorectal cancer via AMPK and PI3K/AKT/mTOR pathways.

Colorectal cancer (CRC) is the third commonly diagnosed malignancy and the second leading cause of cancer death worldwide. Development of novel chemotherapeutics is crucial. Natural products are the main source of drug discovery, and epipolythiodioxopiperazine (ETP) alkaloids are one kind of them have been reported to have potent biological activities. In the present study, we first isolated Chaetocochin J (CJ), an ETP alkaloid from the secondary metabolites of Chaetomium sp, and studied the anti-CRC activity and mechanism of it. The results showed that CJ exhibits potent proliferation inhibition effect, its IC50 to CRC cells are around 0.5 µM. CJ also induces apoptosis of CRC cells in a dose-dependent manner, and this effect is stronger than topotecan. In addition, CJ treatment triggers autophagic flux in CRC cells, inhibition of autophagy by chloroquine didn't affect CJ-induced apoptosis and growth inhibition, suggesting CJ may simultaneously induced apoptosis and autophagy in CRC cells. We further explored the mechanism of action, and found that CJ exerts its anti-CRC function via AMPK and PI3K/AKT/mTOR pathways and further regulation of their downstream signaling cascade in CRC cells, including apoptosis and autophagy. These data potently suggest that CJ may be a potential drug candidate for CRC treatment.

Single-cell transcriptomics reveals intestinal cell heterogeneity and identifies Ep300 as a potential therapeutic target in mice with acute liver failure.

Acute liver failure (ALF) is a severe life-threatening disease associated with the disorder of the gut-liver axis. However, the cellular characteristics of ALF in the gut and related therapeutic targets remain unexplored. Here, we utilized the D-GALN/LPS (D/L)-induced ALF model to characterize 33,216 single-cell transcriptomes and define a mouse ALF intestinal cellular atlas. We found that unique, previously uncharacterized intestinal immune cells, including T cells, B cells, macrophages, and neutrophils, are responsive to ALF, and we identified the transcriptional profiles of these subsets during ALF. We also delineated the heterogeneity of intestinal epithelial cells (IECs) and found that ALF-induced cell cycle arrest in intestinal stem cells and activated specific enterocyte and goblet cell clusters. Notably, the most significantly altered IECs, including enterocytes, intestinal stem cells and goblet cells, had similar activation patterns closely associated with inflammation from intestinal immune activation. Furthermore, our results unveiled a common Ep300-dependent transcriptional program that coordinates IEC activation during ALF, which was confirmed to be universal in different ALF models. Pharmacological inhibition of Ep300 with an inhibitor (SGC-CBP30) inhibited this cell-specific program, confirming that Ep300 is an effective target for alleviating ALF. Mechanistically, Ep300 inhibition restrained inflammation and oxidative stress in the dysregulated cluster of IECs through the P38-JNK pathway and corrected intestinal ecology by regulating intestinal microbial composition and metabolism, thereby protecting IECs and attenuating ALF. These findings confirm that Ep300 is a novel therapeutic target in ALF and pave the way for future pathophysiological studies on ALF.

Asperemestrins A-D, Emestrin Hybrid Polymers with Bridged Skeletons from the Endophytic Fungus Aspergillus nidulans.

Four emestrin hybrid polymers, asperemestrins A-D (1-4, respectively), were isolated from the fungus Aspergillus nidulans. Asperemestrins A-C are the first examples of emestrin-sterigmatocystin heterodimers bearing a 7/5/6/6/5/5/6/6/6 nonacyclic system with a 2,5-diazabicyclo[2.2.2]octane-3,6-dione core, while asperemestrin D features an unprecedented 2,15-dithia-17,19-diazabicyclo[14.2.2]icosa-4,8-diene-12,18,20-trione core skeleton. Their structures were determined by extensive spectroscopic data, electronic circular dichroism calculations, and single-crystal X-ray diffraction. Asperemestrin B showed moderate cytotoxicity against cancer cell lines, including SU-DHL-2, HEPG2, and HL-60.

Lance-Adams syndrome: a report of two cases.

Chronic post-hypoxic myoclonus, also known as Lance-Adams syndrome (LAS), is a rare complication of successful cardiopulmanry resuscitation often accompanied by action myoclonus and cerebellar ataxia. It is seen in patients who have undergone a cardiorespiratory arrest, regained consciousness afterwards, and then developed myoclonus days or weeks after the event. Worldwide, 122 cases have been reported in the literature so far, including 1 case of Chinese. Here we report 2 Chinese LAS patients with detailed neuroimagings. Cranial single photon emission computed tomography (SPECT) of patient 1, a 52-year-old woman, showed a mild hypoperfusion in her left temporal lobe, whereas patient 2, a 54-year-old woman, manifested a mild bilateral decrease of glucose metabolism in the frontal lobes and a mild to moderate decrease of the N-acetyl aspartate (NAA) peak in the bilateral hippocampi by cranial [(18)F]-fluorodeoxyglucose positron emission tomographic (PET) scan and cranial magnetic resonance spectroscopy (MRS), respectively. We also review the literature on the neuroimaging, pathogenesis, and treatment of LAS.

[IT15 gene analysis in two pedigrees of Huntington's disease].

To investigate the relationship between the clinical features and (CAG)n trinucleotide repeats in two pedigrees of Chinese Huntington's disease (HD). Clinical and neuroimaging features, the age of disease onset and pattern of transmission of the patients were studied in the two pedigrees of HD. Genomic DNA of 42 family members was used for amplification of the (CAG)n repeats of IT15 gene by PCR. The numbers of (CAG)n were determined by electrophoresis through a 6% polyacrylamide gel and direct sequence analysis. Results showed that patients in pedigree 1 were absent of the typical triad of HD symptoms or caudate atrophy. A total of 9 (5 patients and 4 asymptomatic) out of 18 family members had 40-50 (CAG)n repeats in the IT15 gene. In pedigree 2, all the patients were characterized by a triad of symptoms, including motor disturbance, cognitive impairment and psychiatric features. Three patients and two asymptomatic relatives had more than 50 (CAG)n repeats in the IT15 gene. In conclusion, the clinical symptoms are partly determined by (CAG)n repeats in the IT15 gene. The age of onset was correlated with (CAG)n repeats over 50, and the phenomenon called "anticipation" was found to have played a role.

GM1 stabilizes expression of NMDA receptor subunit 1 in the ischemic hemisphere of MCAo/reperfusion rat.

OBJECTIVE: To determine the protective effect of monosialoganglionside (GM1) and evaluate the influence of GM1 on expression of N-methyl-D-aspartate receptor subunit 1 (NMDAR1) in Sprague-Dawley (SD) rats with focal cerebral ischemia-reperfusion (I/R). METHODS: Left middle cerebral artery (MCA) was occluded by an intraluminal suture for 1 h and the brain was reperfused for 72 h in SD rats when infarct volume was measured, GM1 (10 mg/kg) was given ip (intraperitoneally) at 5 min (group A), 1 h (group B) and 2 h (group C) after MCA occlusion (MCAo). Expression of NMDAR1 was detected by Western blot at various time after reperfusion (4 h, 6 h, 24 h, 48 h and 72 h) in ischemic hemispheres of the rats with or without GM1 administered. RESULTS: (1) Adjusted relative infarct volumes of groups A and B were significantly smaller than that of group C and the control group (P<0.01 and P<0.05, respectively). (2) Expression level of NMDAR1 was temporally high at 6 h after reperfusion, and dipped below the normal level at 72 h after reperfusion. GM1 at 5 min after MCAo significantly suppressed the expression of NMDAR1 at 6 h after reperfusion (P<0.05 vs the control). At 72 h after reperfusion, the NMDAR1 expression level of rats treated with GM1 administered (at 5 min or 2 h after MCAo) was significantly higher than that of the control (P<0.05). CONCLUSION: GM1 can time-dependently reduce infarct volume in rats with focal cerebral I/R partly through stabilizing the expression of NMDAR1.

[Change of serum soluble intercellular adhesion molecule and basic fibroblast growth factor in patients with acute cerebral infarction and its clinical significance].

OBJECTIVE: To explore the changes of the levels of soluble intercellular adhesion molecule-1 (sICAM-1) and basic fibroblast growth factor (bFGF) in serum of patients with acute cerebral infarction, and the effects of sICAM-1 and bFGF on cerebral infarction. METHODS: ELISA was used to detect the serum sICAM-1 and bFGF of 55 patients with acute cerebral infarction (within 2 days) as well as 32 patients diagnosed as with other neurologic diseases (20 patients with sciatica and 12 with trigeminal neuralgia) and 30 healthy persons as controls. RESULTS: (1) Both serum sICAM-1 and bFGF in the infarction group were significantly higher [(766.2 +/- 178.8) micro g/L and (17.4 +/- 8.2) micro g/L respectively] than in other disease control group [(529.6 +/- 76.7) micro g/L and (8.3 +/- 2.8) micro g/L respectively] and normal control group [(520.7 +/- 115.9) micro g/L and (5.8 +/- 2.7) micro g/L respectively] (P = 0.000). (2) Correlation analysis showed that there was a positive correlation between the level of serum sICAM-1 and bFGF (r = 0.471, P = 0.000), the level of sICAM-1 was also positively correlated with the number of leukocytes at acute stage (r = 0.285, P = 0.035), and a negative correlation between sICAM-1 and European stroke scale (r = -0.333, P = 0.013) was found. No significant correlation was observed between the level of sICAM-1 and the levels of serum cholesterol (r = -0.042, P = 0.758) or triglyceride (r = 0.061, P = 0.675). (3) Blood pressure seemed to have no influence on the content of sICAM-1 and bFGF after cerebral infarction, while the level of serum bFGF with large infarct size was obviously higher at acute stage. CONCLUSION: The levels of sICAM-1 and bFGF increase significantly in the patients with acute cerebral infarction. sICAM-1 and bFGF mayin participate in the pathophysiologic process through inflammatory mechanism. The detection of serum sICAM-1 will be helpful in estimating the clinical severity and the determination of bFGF will be helpful in estimating the size of infarct lesion at acute stage of cerebral infarction.

[Expression of urokinase-type plasminogen activator and its receptor in plasma of patients with cerebral infarction].

OBJECTIVE: To measure the urokinase-type plasminogen activator (UPA) and its receptor (UPAR) in the plasma of patients with cerebral infarction and to study the effects of UPA and UPAR on cerebral infarction. METHODS: ELISA sandwich method was used to measure the plasma levels of UPA and UPAR in 89 patients with acute cerebral infarction, subdivided into mild, moderate and severe subgroups according to the modified Edinburgh-Scandinavia stroke scale (mESSS), and 30 patients with other disease and 30 healthy persons as controls. RESULTS: Within 2 days after the onset of cerebral infarction the UPA and UPAR levels were (1663.7 +/- 384.2) ng/L and (1 375.3 +/- 303.0) ng/L respectively, both significantly higher than those of the patients with other diseases [(1 033.0 +/- 122.7) ng/L and (978.3 +/- 120.0) ng/L respectively (P = 0.038 and P = 0.000)] and those of the normal control group [(1 005.0 +/- 128.9) ng/L and (904.7 +/- 158.6) ng/L respectively, both P = 0.000]. Two weeks after the onset of stroke, the plasma UPA level was (1 185.5 +/- 384.6) ng/L, not significantly different from those of the 2 control groups (both P > 0.05); while the plasma UPAR level in the cerebral infarction was (1 159.3 +/- 261.2) ng/L, significantly higher than those in the 2 control groups (P < 0.01). Within 2 days after the onset of infarction the plasma UPA and UPAR levels of the severe subgroup were (1 938.9 +/- 256.5) ng/L and (1 510.8 +/- 378.7) ng/L respectively, both significantly higher than those of the moderate subgroup [(1 593.7 +/- 204.8) ng/L and (1 296.6 +/- 151.2) ng/L respectively, both P < 0.01] and those of the mild subgroup [1 358.5 +/- 175.9] ng/L and (1 226.8 +/- 98.3) ng/L respectively, both P < 0.01], Two weeks after the onset of stroke,the plasma UPA and uPAR levels of the severe subgroup remained significantly higher than those of the mild subgroup [(1 152.6 +/- 170.3) ng/L vs (1 041.9 +/- 187.0) ng/L, (P < 0.05)] and [(1 186.4 +/- 158.3) ng/L vs (1 053.9 +/- 109.4) ng/L, (P < 0.01)]. In addition, the plasma UPA and UPAR levels of the patients who died from cerebral infarction at last were the highest among all the groups. CONCLUSION: The plasma levels pf UPA and UPAR increase in patients with cerebral infarction and may be related with the severity of disease. The UPA and UPAR genes may play an important role via proteolytic degradation of the extracellular matrix and basement membrane during the development of cerebral infarction.

[Protective effects and mechanisms of serial TCM "Huoxuehuayu" prescriptions on cerebral ischemia-reperfusion injury in rats].

OBJECTIVE: To determine effects and mechanisms of traditional Chinese medicine serial "Huoxuehuayu" (activating blood flow and removing blood stasis) prescriptions on cerebral ischemia-reperfusion injury in rats. METHODS: Focal cerebral ischemia was induced by 60 min of middle cerebral artery occlusion (MCAO) and followed by 3 d reperfusion. Drugs were orally administered 1 h before MCAO and 4 h after reperfusion and the following 2 d. The neurological scores were evaluated 3 d after reperfusion. Then the animals were sacrificed, the infarct volumes, right and left areas of brain section, the pathologic changes and the normal neurons in hippocampal CA1 and cortex were determined by using an image analyzer. Nitric oxide synthase (NOS), superoxide dismutase (SOD) activities and malondialdehyde (MDA) content in brain tissue were evaluated by spectrophotography. The expression of NMDA R1 subunit (NR1) and endothelial NOS (eNOS) was determined by immunoblotting technique. RESULT: Serial "Huoxuehuayu" prescriptions and nimodipine improved neuronal dysfunction, and reduced infarct size and brain edema. Serial Huoxuehuayu prescriptions and nimodipine reduced MDA content and NOS activity, and increased SOD activity. Western blotting analysis demonstrated induction of NR1. CONCLUSION: Serial Huoxuehuayu prescriptions have a protective effect on cerebral ischemia-reperfusion injury by reducing NOS activity, MDA content, expression of NR1 and increasing SOD activity.

[Serum levels of soluble intercellular adhesion molecule-1 in patients with cerebral infarct].

OBJECTIVE: To observe the changes of the levels of soluble intercellular adhesion molecule-1(sICAM-1) in serum of patients with cerebral infarct and to explore the effect of sICAM-1 on cerebral infarct. METHODS: The serum levels of sICAM-1 in 55 patients with cerebral infarct both in acute stage(within 2 days) and convalescence(2 weeks after attack) were detected by using ELISA. At the same time, we compare the results with those of 32 patients having other neurologic diseases(20 patients with sciatica, 12 with trigeminal neuralgia) and 30 healthy subjects. RESULTS: (1) The serum levels of sICAM-1 of patients with cerebral infarct (acute stage: 766+/-179 microgram/L, convalescence: 602+/-155 microgram/L, respectively) were significantly higher than those of the control groups(530+/-77 microgram/L and 521+/-116 microgram/L, respectively, P<0.01). (2)There was a positive correlation of SICAM levels with the amount of leukocytes in acute stage(r=0.285,P<0.05), but negative correlation to clinical severity of cerebral infarct(r= 0.333,P<0.05). And there was no significant correlation between the level of sICAM-1 and the levels of cholesterol and triglyceride in serum(r= 0.042 and r=0.061, respectively, P>0.05). (3)There was no significant difference between sICAM levels of patients of cerebral cortex infarct and those of patients with basal ganglia infarct(773+/-178 microgram/L and 758+/-183 microgram/L, respectively, P>0.05). (4)The levels of sICAM-1 between patients of cerebral infarct with or without hypertension were no significant difference(774+/-189 microgram/L and 754+/-165 microgram/L, respectively, P>0.05). CONCLUSION: The levels of sICAM-1 increase significantly in patients with cerebral infarct. sICAM-1 may participate in the pathophysiologic process through inflammatory mechanism.

[Study of brainstem auditory evoked potentials and its correlation with pontine volume in olivopontocerebellar atrophy].

AIM: To investigate the change of latency and interpeak latency of each component of BAEP (brainstem auditory evoked potential, BAEP) and its correlation with PV/PFV (pontine volume/posterior fossa volume, PV/PFV) ratio in OPCA (olivopontocerebellar atrophy, OPCA). METHODS: We used Keypoint EMG/EP to determine waves I PL (peak latency, PL), III PL, V PL and I - III IPL (interpeak latency, IPL), III - V IPL, I - V IPL and used 1.5TMR 3D volume rendering software to determine PV (pontine volume, PV), CV(cerebellar volume, CV) and PFV (posterior fossa volume,PFV). Then calculated PV/PFV ratio, CV/PFV ratio and PV/ CV ratio in OPCA group and control group. RESULTS: Compared with control group, in OPCA group wave IIII PL, I - III IPL were significantly elongated (P < 0.05), III - V IPL was significantly shorten (P < 0.05), PV/PFV ratio was significantly decreased (P < 0.01); there was a positive correlation between III-V IPL and PV/PFV ratio (r = 0.83, P < 0.01). CONCLUSION: In patients with OPCA, III PL, I - III IPL of BAEP were elongated and III - V IPL of BAEP was shorten. III - V IPL became shorter when the volume of pontine decreased.

Medullary hemorrhagic infarction after radiation for nasopharyngeal carcinoma.

Head and neck irradiation may lead to accelerated atherosclerosis over several years. Delayed stroke has been described after head and neck irradiation administered for a number of conditions. However, brain stem stroke has only rarely been associated with irradiation. We report a patient with medullary hemorrhagic infarction 6 years after radiotherapy for nasopharyngeal carcinoma. A 42-year-old normotensive Chinese male had rapid onset of vertigo, diplopia, ataxia, dysphagia, hypophonic dysarthria, hemiparesis, and respiratory distress. Cranial MR imaging 2 days after symptom onset showed medullary infarction, and cranial MR imaging 5 days after symptom onset showed medullary hemorrhage. He needed ventilatory support and died of bacterial pneumonia 1 month later. Other risk factors for stroke were absent. Hemorrhagic infarction in this patient was likely associated with the radiotherapy. Radiotherapy is the first choice of treatment for nasopharyngeal carcinoma, however, it may induce fatal medullary hemorrhagic infarction.

Monosialoganglioside protected ischemic rat hippocampal slices through stabilizing expression of N-methyl-D-aspartate receptor subunit.

AIM: To determine direct protective effect of monosialoganglioside (GM1) on hippocampal slices after oxygen-glucose deprivation and reperfusion (OGD/RP), and investigate the influence on the expression of N-methyl-D-aspartate receptor subunit 1 (NMDAR1) in those hippocampal slices. METHODS: Injury of hippocampal slices and protective effects of GM1 were detected by 2,3,5-triphenyltetrazolium chloride (TTC) staining, toluidine blue staining, and transmission electron microscopy of rat hippocampal slices. Expression of NMDAR1 was detected by Western blot. RESULTS: (1) GM1 at 1.0 micromol/L was the most effective concentration to preserve the TTC staining of the hippocampal slices after OGD/RP (P<0.05), and the next was GM1 at 10.0 micromol/L (P<0.05). (2) Toluidine blue staining and transmission electron microscopy showed GM1 protected the injuried hippocamal slices after OGD/RP. (3) GM1 downregulated the temporally high expression of NMDAR1 in the hippocampal slices immediately after a 25-min OGD and prevented the over low expression of NMDAR1 after a 30-min reperfusion. CONCLUSION: GM1 could protect injuried rat hippocampal slices after OGD/RP through stabilizing the expression of NMDAR1.