[The clinical characteristics of neutrophil extracellular trap in patients with severe cerebral venous sinus thrombosis and its prognostic value in acute and subacute patients].

Objective: To explore the clinical characteristics of neutrophil extracellular trap (NET) in patients with severe cerebral venous sinus thrombosis (CVST) and to study their prognostic value in the acute and subacute phases. Methods: This study is a retrospective case series analysis. Clinical and pathological data of 52 patients with severe cerebral venous sinus thrombosis who underwent endovascular treatment in the Department of Neurosurgery, Tianjin Huanhu Hospital from June 2019 to June 2022 were retrospectively analyzed. There were 20 males and 32 females, with an age of (40.1±13.6) years(range:18 to 66 years). Forty-five healthy physical examinees were included in the control group. High-resolution MRI was used to stage the thrombus, with 11 cases in the acute group, 28 cases in the subacute group, and 13 cases in the chronic group. Thrombus specimens were obtained through endovascular treatment, and the fluorescence intensity of NET in peripheral blood at different time points was analyzed by immunofluorescence contrast,including the double-stranded DNA structure and adhesion protein components (citrolinated histone H3 (CitH3), myeloperoxidase-DNA complex(MPO-DNA), neutrophil elastase (NE)). The NET markers were determined by ELISA. Spearman rank correlation analysis was used to analyze the correlation between the NET markers in peripheral blood of patients with severe cerebral venous sinus thrombosis in the acute and subacute phases and the volume of venous sinus thrombus, the degree of venous sinus recanalization after treatment, and the discharge modified Rankin scale(mRS)score. The accuracy of NET markers in predicting the prognosis of patients with severe cerebral venous sinus thrombosis was analyzed by drawing receiver operating characteristic (ROC) curve and calculating the area under the curve (AUC). Results: The results of immunofluorescence staining and ELISA showed that no NET structure was formed in the peripheral blood of the control group, while CitH3, MPO-DNA and NE levels in the peripheral blood of CVST patients were increased, among which the acute stage group was the highest, followed by the subacute group, and the chronic group was the lowest. Spearman correlation analysis showed that CitH3, MPO-DNA and NE levels in peripheral blood of patients in acute group and subacute group were positively correlated with thrombus volume and mRS score at discharge (P<0.05). The levels of CitH3 and MPO-DNA in peripheral blood of patients with complete venous sinus recanalization were lower than those of patients with partial venous sinus recanalization (P<0.01). ROC curve analysis results showed that MPO-DNA and NE had no predictive ability for the prognosis of CVST patients (P values were 0.614 and 0.324, respectively), and the AUC of CitH3 was 0.800 (95%CI: 0.638~0.962, P=0.032), the best cut-off value was 13.5 µg/L, the sensitivity was 100%, and the specificity was 58.8%. Conclusions: A large number of NET are formed in patients with severe cerebral venous sinus thrombosis in acute stage. Patients with severe cerebral venous sinus thrombosis in acute stage and subacute stage with high peripheral blood NET content has a low rate of complete sinus revascularization and poor neurological function recovery after treatment.

Engineering tumor-colonizing E. coli Nissle 1917 for detection and treatment of colorectal neoplasia.

Bioengineered probiotics enable new opportunities to improve colorectal cancer (CRC) screening, prevention and treatment. Here, first, we demonstrate selective colonization of colorectal adenomas after oral delivery of probiotic E. coli Nissle 1917 (EcN) to a genetically-engineered murine model of CRC predisposition and orthotopic models of CRC. We next undertake an interventional, double-blind, dual-centre, prospective clinical trial, in which CRC patients take either placebo or EcN for two weeks prior to resection of neoplastic and adjacent normal colorectal tissue (ACTRN12619000210178). We detect enrichment of EcN in tumor samples over normal tissue from probiotic-treated patients (primary outcome of the trial). Next, we develop early CRC intervention strategies. To detect lesions, we engineer EcN to produce a small molecule, salicylate. Oral delivery of this strain results in increased levels of salicylate in the urine of adenoma-bearing mice, in comparison to healthy controls. To assess therapeutic potential, we engineer EcN to locally release a cytokine, GM-CSF, and blocking nanobodies against PD-L1 and CTLA-4 at the neoplastic site, and demonstrate that oral delivery of this strain reduces adenoma burden by ~50%. Together, these results support the use of EcN as an orally-deliverable platform to detect disease and treat CRC through the production of screening and therapeutic molecules.

[Long-term efficacy of balloon-assisted endplate augmentation combined with transforaminal pedicle screw fixation in the treatment of thoracolumbar burst fractures].

Objectives: To investigate the long-term efficacy of balloon assisted endplate reduction with vertebral augmentation combined with pedicle screw fixation in the treatment of thoracolumbar burst fractures, and to compare the clinical efficacy of calcium sulfate cement (CSC) and calcium phosphate cement(CPC). Methods: This study is a retrospective cohort study.The clinical data of 39 patients with thoracolumbar burst fractures admitted to Hefei Hospital Affiliated to Anhui Medical University from November 2013 to December 2017 were retrospectively analyzed.All patients were treated with pedicle screw reduction and fixation of the injured vertebra,balloon-assisted reduction of the collapsed endplate of the injured vertebra,and artificial bone vertebral body augmentation,and the follow-up time was >5 years.There were 24 males and 15 females,aged (42.9±13.3) years (range: 29 to 56 years).According to the Frankel spinal nerve dysfunction grading standard, there were 4 cases of grade C, 7 cases of grade D and 28 cases of grade E. There were 21 cases of CSC augmentation(CSC group) and 18 cases of CPC augmentation (CPC group). X-ray and CT were performed at 1 week, 1-, 2-, 5-year after surgery and at the last follow-up, and the imaging indicators were measured, including the injured vertebra anterior edge height ratio,the injured vertebra middle height ratio,the injured vertebra wedge angle,and the sagittal plane Cobb angle. The pain visual analogue scale (VAS) and the Oswestry disability index (ODI) was used for functional evaluation, nervous function was evaluated according to the Frankel spinal nerve dysfunction grading standard.Independent sample t test was used for inter-group comparison, and paired sample t test and repeated measure ANOVA were used for intra-group comparison. Results: All operative procedures were successfully completed, no spinal nerve function damage occurred. The postoperative imaging indexes of the patients were significantly improved compared with those before surgery (all P<0.01). The follow-up time of patients was (6.7±2.8)years (range: 5 to 9 years). Among the 11 patients with symptoms of neurological impairment before surgery, 9 patients completely recovered at the last follow-up, and 2 patients recovered from Frankel grade C to D. There were no significant differences in imaging indexes between the first week after surgery and the last follow-up in the CPC group (all P>0.05), while there were significant differences in imaging indexes between the CSC group and the last follow-up (all P<0.05). CPC group was superior to CSC group in frontal height ratio, middle height ratio, wedge angle variation and sagittal Cobb angle correction loss at 2 year, 5 year after surgery and the last follow-up, with statistical significance (all P<0.05). At the last follow-up, there were no differences in VAS and ODI between the two groups (all P>0.05). After absorption of CSC in the filling area, a hardened zone was formed around the area, and the central cavity remained without bone tissue filling. CPC absorption was very slow, and the CPC group was still filled satisfactorily at the last follow-up. Conclusions: Balloon assisted endplate reduction and vertebral augmentation combined with pedicle screw fixation through the injured vertebra have good long-term efficacy in the treatment of thoracolumbar burst fractures. Compared with CSC, CPC vertebral augmentation can better maintain the shape and spinal sequence of the injured vertebra in the long term, and can effectively reduce the collapse of the space above the injured vertebra.

[Intervention effect of apocynin on silicosis induced by silica in rats].

Objective: To investigate the intervention effect and its mechanism of apocynin, an inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) on silicosis induced by silica (SiO(2)) in rats. Methods: In October 2021, 24 SPF SD male rats were divided into control group, silicosis model group and apocynin intervention group according to random number table method, with 8 rats in each group. SiO(2) was exposed by one-time intratracheal instillation. The rats in the apocynin intervention group were intraperitoneally injected with apocynin 50 mg/kg, 3 times a week, on the second day after treatment. The rats were sacrificed 28 days later, and lung coefficients were calculated after lung tissues were weighed. Hematoxylin-eosin staining and Masson staining were used to observe the lung histopathological changes in each group, respectively. The levels of NOX, reactive oxygen species (ROS), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in lung tissue were detected. The expressions of interleukin-1 beta (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined by Enzyme-Linked Immunosorbent Assay (ELISA). The level of hydroxyproline (HYP) was detected by alkaline hydrolysate. The expressions of transforming growth factor beta 1 (TGF-ß1), E-cadherin (E-cad) and α-smooth muscle actin (α-SMA) in lung tissue were detected by Western blotting. Results: Compared with the control group, the body weight of silicosis model group was decreased, the lung tissue showed obvious inflammatory infiltration and fibrosis, and the levels of lung coefficient, IL-1ß, IL-6, TNF-α and TGF-ß1 were significantly increased (P<0.05). Compared with the silicosis model group, the lung tissue injury in the apocynin intervention group was significantly improved, the lung coefficient, NOX, ROS, MDA, IL-1ß, IL-6, TNF-α and TGF-ß1 levels were decreased, and the activity of GSH-Px was increased (P<0.05). Compared with the silicosis model group, the expressions of HYP and α-SMA were decreased and the level of E-cad was increased in the apocynin intervention group (P<0.05) . Conclusion: Apocynin may alleviate SiO(2)-induced fibrosis in silicosis rats by reducing oxidative stress, the release of inflammatory factors and inhibiting the process of epithelial-mesenchymal transition.

[The value of aspirin challenge tests in the diagnosis of non-steroidal anti-inflammatory drugs-exacerbated respiratory disease].

Objective: To investigate the value of aspirin challenge tests in the diagnosis of non-steroidal anti-inflammatory drugs-exacerbated respiratory disease (NERD). Methods: Fifty patients (22 males and 28 females; aged 16-61 years) who were diagnosed with chronic rhinosinusitis with nasal polyps (CRSwNP) with/without asthma, and underwent NERD standardized diagnosis in the Allergy Centre of West China Hospital, Sichuan University from December 2021 to November 2022 were included in the study. The first step was asking about the history of exacerbation respiratory symptoms after intake of any non-steroidal anti-inflammatory drug, including aspirin; the second step was performing intranasal aspirin challenge (IAC); and the third step was performing oral aspirin challenge (OAC). The diagnosis of NERD was made if any of the above steps was positive, and the subsequent steps were not performed, otherwise the diagnosis was made to OAC. If OAC was negative, the diagnosis was non-NERD. All patients completed the sino-nasal outcome test 22 (SNOT 22) score, Lund-Kennedy score by nasal endoscopic, allergen skin prick test, blood routine and serum total IgE test. SPSS version 20.0 was used for statistical analysis. Results: The diagnosis of NRED was confirmed in 27 patients (27/50, 54%). Seven (7/50, 14%) of them were diagnosed by clinical history and 20 (20/50, 40%) were diagnosed by aspirin challenge tests, of which 17 (17/20, 85%) were positive to IAC and 3 (3/20, 15%) to OAC. Of the 43 patients who underwent IAC testing, only 2 (2/43, 5%) developed asthma attacks during challenge. Comparing the clinical characteristics of patients in NERD and non-NERD group, there were significant differences between the two groups in gender (P=0.001), hyposmia (P=0.003), history of repeated CRSwNP surgeries (P=0.028), comorbid asthma (P=0.013), SNOT-22 score (P=0.004) and the percentage of peripheral blood eosinophil (P=0.043). Conclusions: Patients may be underdiagnosed if the diagnosis of NERD is made only by medical history, and it is necessary to carry out aspirin challenge tests. IAC is an important means to diagnose NERD with high accuracy and good safety. However, If IAC is negative, further OAC is required.

Engineered bacteria detect tumor DNA.

Synthetic biology has developed sophisticated cellular biosensors to detect and respond to human disease. However, biosensors have not yet been engineered to detect specific extracellular DNA sequences and mutations. Here, we engineered naturally competent Acinetobacter baylyi to detect donor DNA from the genomes of colorectal cancer (CRC) cells, organoids, and tumors. We characterized the functionality of the biosensors in vitro with coculture assays and then validated them in vivo with sensor bacteria delivered to mice harboring colorectal tumors. We observed horizontal gene transfer from the tumor to the sensor bacteria in our mouse model of CRC. This cellular assay for targeted, CRISPR-discriminated horizontal gene transfer (CATCH) enables the biodetection of specific cell-free DNA.

Baicalin alleviates silica-induced lung inflammation and fibrosis by inhibiting TLR4/NF-?B pathway in rats.

Silicosis is an occupational lung disease caused by inhaling silica dust. The disease is characterized by early lung inflammation and late irreversible pulmonary fibrosis. Here we report the effect of Baicalin, a main flavonoid compound from the roots of Chinese herbal medicine Huang Qin on silicosis in a rat model. Results showed Baicalin (50 or 100 mg/kg/day) can mitigate the silica-induced lung inflammation and reduce the harm of alveolar structure and the blue region of collagen fibers in rat lung at 28 days after administration. At the same time, Baicalin also diminished the level of interleukin-1beta (IL-1beta, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) in lung tissues. The protein expression of collagen I (Col-1), alpha-smooth muscle actin (alpha-SMA) and vimentin were down-regulated while E-cadherin (E-cad) was increased in Baicalin-treated rats. In addition, the Toll Like Receptor 4 (TLR4)/ nuclear factor kappaB (NF-kappaB) pathway was enabled at 28 days after silica infusion, and the treatment of Baicalin diminished the expression of TLR4 and NF-?B in the lungs of rat with silicosis. These results suggested that Baicalin inhibited the pulmonary inflammatory and fibrosis in a rat model of silicosis, which could be attributed to inhibition of the TLR4/NF-kappaB pathway.

Colorectal cancer detection and treatment with engineered probiotics.

Bioengineered probiotics enable new opportunities to improve colorectal cancer (CRC) screening, prevention and treatment strategies. Here, we demonstrate the phenomenon of selective, long-term colonization of colorectal adenomas after oral delivery of probiotic E. coli Nissle 1917 (EcN) to a genetically-engineered murine model of CRC predisposition. We show that, after oral administration, adenomas can be monitored over time by recovering EcN from stool. We also demonstrate specific colonization of EcN to solitary neoplastic lesions in an orthotopic murine model of CRC. We then exploit this neoplasia-homing property of EcN to develop early CRC intervention strategies. To detect lesions, we engineer EcN to produce a small molecule, salicylate, and demonstrate that oral delivery of this strain results in significantly increased levels of salicylate in the urine of adenoma-bearing mice, in comparison to healthy controls. We also assess EcN engineered to locally release immunotherapeutics at the neoplastic site. Oral delivery to mice bearing adenomas, reduced adenoma burden by ∻50%, with notable differences in the spatial distribution of T cell populations within diseased and healthy intestinal tissue, suggesting local induction of robust anti-tumor immunity. Together, these results support the use of EcN as an orally-delivered platform to detect disease and treat CRC through its production of screening and therapeutic molecules.

[Study of risk factors of postherpetic neuralgia].

Objective: To analyze the clinical features and risk factors of herpes zoster and the risk factors of postherpetic neuralgia (PHN). Methods: A total of 2 840 patients diagnosed with herpes zoster in Peking University People's Hospital from January 2021 to April 2022 were included. The patients with herpes zoster were aged (59±16), and of which 1 314 (46.3%) patients were male. The patients were divided into PHN group and non-PHN group according to whether the patients with herpes zoster developed PHN. And there were 442 (15.6%) patients developed PHN aged (68±12), and of which 189 (42.8%) were male. The information and medical history of PHN group and non-PHN group were collected. The information of the use of glucocorticoids, immunosuppressive agents, biological agents and targeted medicine within 2 months before herpes zoster occurred, history of surgery within 6 months before herpes zoster occurred, and the duration of PHN were collected. Logistic regression analysis was used to analyze the risk factors of PHN. Results: Out of 2 840 patients, 2 056 (72.4%) with herpes zoster aged 50 years and above. Intercostal nerves was mostly involved in patients with herpes zoster, with a total of 1 532(53.9%). Hypertension (465 cases, 16.4%) accounted for the highest number of patients with chronic diseases, followed by diabetes (337 cases, 11.9%) and coronary heart disease (283 cases, 10.0%). Rheumatoid arthritis (41 cases, 1.4%) accounted for the highest number of patients with connective tissue diseases, followed by Sjogren's syndrome (31 cases, 1.1%), systemic lupus erythematosus (28 cases, 1.0%). Logistic regression analysis showed that age≥50 years old (OR: 4.581; 95%CI: 3.131-6.705),lesion on the upper limb and shoulder (OR: 1.858; 95%CI: 1.129-3.059), hypertension (OR: 1.963; 95%CI: 1.513-2.546) and immunosuppressive treatments (OR: 2.170; 95%CI: 1.254-3.753) were independent risk factors for PHN (all P<0.05). Conclusions: Herpes zoster mostly occurs in people aged 50 and above, and mainly affected intercostal nerves. The most common complications are hypertension and rheumatoid arthritis. Age≥50, lesion on upper limbs and shoulders, hypertension, and immunosuppressive treatments may be independent risk factors of PHN.

Structural Analysis of the Complex of Human Transthyretin with 3',5'-Dichlorophenylanthranilic Acid at 1.5 Å Resolution.

Human transthyretin (hTTR) can form amyloid deposits that accumulate in nerves and organs, disrupting cellular function. Molecules such as tafamidis that bind to and stabilize the TTR tetramer can reduce such amyloid formation. Here, we studied the interaction of VCP-6 (2-((3,5-dichlorophenyl)amino)benzoic acid) with hTTR. VCP-6 binds to hTTR with 5 times the affinity of the cognate ligand, thyroxine (T4). The structure of the hTTR:VCP-6 complex was determined by X-ray crystallography at 1.52 Šresolution. VCP-6 binds deeper in the binding channel than T4 with the 3',5'-dichlorophenyl ring binding in the 'forward' mode towards the channel centre. The dichlorophenyl ring lies along the 2-fold axis coincident with the channel centre, while the 2-carboxylatephenylamine ring of VCP-6 is symmetrically displaced from the 2-fold axis, allowing the 2-carboxylate group to form a tight intermolecular hydrogen bond with Nζ of Lys15 and an intramolecular hydrogen bond with the amine of VCP-6, stabilizing its conformation and explaining the greater affinity of VCP-6 compared to T4. This arrangement maintains optimal halogen bonding interactions in the binding sites, via chlorine atoms rather than iodine of the thyroid hormone, thereby explaining why the dichloro substitution pattern is a stronger binder than either the diiodo or dibromo analogues.

The Origin and Contribution of Cancer-Associated Fibroblasts in Colorectal Carcinogenesis.

BACKGROUND & AIMS: Cancer-associated fibroblasts (CAFs) play an important role in colorectal cancer (CRC) progression and predict poor prognosis in CRC patients. However, the cellular origins of CAFs remain unknown, making it challenging to therapeutically target these cells. Here, we aimed to identify the origins and contribution of colorectal CAFs associated with poor prognosis. METHODS: To elucidate CAF origins, we used a colitis-associated CRC mouse model in 5 different fate-mapping mouse lines with 5-bromodeoxyuridine dosing. RNA sequencing of fluorescence-activated cell sorting-purified CRC CAFs was performed to identify a potential therapeutic target in CAFs. To examine the prognostic significance of the stromal target, CRC patient RNA sequencing data and tissue microarray were used. CRC organoids were injected into the colons of knockout mice to assess the mechanism by which the stromal gene contributes to colorectal tumorigenesis. RESULTS: Our lineage-tracing studies revealed that in CRC, many ACTA2+ CAFs emerge through proliferation from intestinal pericryptal leptin receptor (Lepr)+ cells. These Lepr-lineage CAFs, in turn, express melanoma cell adhesion molecule (MCAM), a CRC stroma-specific marker that we identified with the use of RNA sequencing. High MCAM expression induced by transforming growth factor ß was inversely associated with patient survival in human CRC. In mice, stromal Mcam knockout attenuated orthotopically injected colorectal tumoroid growth and improved survival through decreased tumor-associated macrophage recruitment. Mechanistically, fibroblast MCAM interacted with interleukin-1 receptor 1 to augment nuclear factor κB-IL34/CCL8 signaling that promotes macrophage chemotaxis. CONCLUSIONS: In colorectal carcinogenesis, pericryptal Lepr-lineage cells proliferate to generate MCAM+ CAFs that shape the tumor-promoting immune microenvironment. Preventing the expansion/differentiation of Lepr-lineage CAFs or inhibiting MCAM activity could be effective therapeutic approaches for CRC.

[Interventional effect of metformin on pulmonary inflammation and pulmonary fibrosis in silicotic rats].

Objective: To investigate the interventional effect of metformin on pulmonary inflammation and pulmonary fibrosis in silicotic rats. Methods: In April 2019, 48 Wistar male rats of SPF grade were randomly divided into negative control group, metformin control group, silicon dioxide (SiO2) model group, low, medium and high dose metformin intervention group according to the random number table method, 8 rats in each group. The SiO2 model group and the low, medium and high dose metformin intervention groups were given 1 ml 50 mg/ml of SiO2 by intratracheal instillation, the negative control group and the metformin control group were given 1 ml normal saline by intratracheal instillation. 24 hours later, the low, medium and high dose metformin intervention groups and the metformin control group were treated with 100, 200, 400 and 400 mg/kg metformin daily, the control and SiO2 model groups received normal saline daily. Then the rats were sacrificed at the 28th day after SiO2 exposure. The changes of rat body weight and pathological examination of rat lung tissue were observed, and the lung organ coefficient, the content of hydroxyproline (HYP) , the expression levels of inflammatory factors transforming growth factor beta1 (TGF-ß1) , tumor necrosis factor-alpha (TNF-α) , interleukin-1beta (IL-1ß) and the protein expression of E-cadherin (E-Cad) , Vimentin, α-SMA were detected. Results: Compared with the negative control group, SiO2 model group had a significant decrease in the body weight of rats (P<0.05) , lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-ß1, TNF-α, IL-1ß were all significantly increased (P<0.05) . Compared with the SiO2 model group, the weights of the rats in the medium and high dose intervention group of metformin increased significantly (P<0.05) . And after intervention with different doses of metformin, the lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-ß1, TNF-α and IL-1ß were significantly decreased (P<0.05) . Immunohistochemistry and Western blotting results showed that compared with the negative control group, the expression of E-Cad of the SiO2 model group was decreased, and the expression levels of Vimentin and α-SMA were significantly increased (P<0.05) . After metformin intervention, the expression of E-Cad was significantly increased, the expression levels of Vimentin and α-SMA were significantly decreased (P<0.05) . Conclusion: Metformin can reduce lung tissue inflammation and fibrosis in rats exposed to SiO2 dust, which may be related to reducing the expression of inflammatory factors in lung tissue and inhibiting the EMT process.

[Evaluation of papilledema and visual improvement in patients with idiopathic intracranial hypertension after venous sinus stenting].

Objective: To evaluate the improvement of papilledema and visual acuities in patients with idiopathic intracranial hypertension (IIH) after venous sinus stenting. Methods: The clinical data of 8 IIH patients who met the inclusion criteria underwent venous sinus stenting between January 2013 and December 2016 at Department of Neurosurgery, Tianjin Huanhu Hospital were analyzed retrospectively. There were 6 females and 3 males,aged (32.9±14.4)years (range:19 to 57 years).The thickness of the retinal nerve fiber layer (RNFL) was measured by optical coherence tomography. Fundus,visual acuity and visual field examination were performed before and after operation. If pressure gradient ≥10 mmHg(1 mmHg=0.133 kPa) across the venous stenosis was indicated by intraoperative pressure measurement,the patient would be treated with venous sinus stenting. Intracranial pressure was measured by lumbar puncture 3 to 7 days after operation. RNFL thickness and eye examination were detected 6 months after surgery. CT venogram was used to observe the sinus venous conditions. Paired t test was used to compare the data before and after surgery. Results: All the 8 patients underwent venous sinus stenting successfully. The mean pressure gradient across the venous stenosis was reduced from (24±9.2) mmHg to (2.6±2.0) mmHg (t=8.02,P<0.01). Intracranial pressure decreased from preoperative (41.4±12.7) cmH2O(1 cmH2O=0.098 kPa) to postoperative (12.9±3.3) cmH2O (t=7.08, P<0.01). The RNFL thickness decreased from (275.3±68.3)µm to (131.4±31.8)µm(t=5.80,P<0.05) 6 months after surgery and the baseline visual acuity was improved from(M(QR))0.24 (0.25) to 0.65 (0.23)(Z=-2.52,P<0.05).Papilledema was significantly improved in 6 patients,and no significant change in 2 patients. CT venogram indicated adjacent stent restenosis in 1 patient. Conclusion: Venous sinus stenting can effectively improve papilledema and visual acuity caused by IIH.

C1q/tumor necrosis factor-related protein-3 acts as a target treating hepatic fibrosis.

OBJECTIVE: C1q/tumor necrosis factor-related protein-3 (CTRP3) is demonstrated as a crucial factor that participated in various fibrotic diseases. Activation of hepatic stellate cell in liver takes a critical effect on the pathogenesis of hepatic fibrosis. However, the role of CTRP3 in hepatic fibrosis remains elusive. Our present study aimed to explore the molecular mechanism of CTRP3 in fibroblast activation and the development of hepatic fibrosis. MATERIALS AND METHODS: We carried out overexpression (OE) of CTRP3 or knockout (KO) of CTRP3 in hepatic stellate cells (HSCs), respectively. Then, transforming growth factor-beta (TGF-ß) was used to stimulate HSCs activation. Adult male C57BL/6J mice were treated tetrachloromethane by intraperitoneal injection and mice injected saline were served as control. Recombinant CTRP3 (RC-CTRP3) was employed to treat CCl4-induced liver fibrosis. Then, the expression of fibrotic biomarkers, Notch signaling pathway-associated factors, liver histology and liver function were investigated in vivo, respectively. RESULTS: Our results showed that CTRP3 decreased in fibrotic liver and TGF-ß treated HSCs. In vitro, CTRP3 inhibited the activation of HSCs and impeded extracellular matrix (ECM) including collagen I and fibronectin via inhibiting Notch-1/Jagged-1 signaling pathway. In vivo, the indexes of fibrogenesis in liver fibrotic mice received RC-CTRP3 were mitigated via regulation of Notch-1/Jagged-1 signaling pathway. Moreover, liver histology and liver function were improved through the increase of CTRP3 level. CONCLUSIONS: The results proved that CTRP3 as a distinguished anti-fibrotic target inhibited HSCs activation by TGF-ß inducement and protected the liver tissue in the process of liver fibrosis.

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma.

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

[Identification of serum biomarkers and evaluation of metabolism disorders in patients with oral squamous cell carcinoma].

Objective: To explore the changes in serum metabolic profile in patients with oral squamous cell carcinoma (OSCC) and to identify the diagnostic biomarkers in order to provide new ideas for the early diagnosis of OSCC. Methods: In total, 76 OSCC patients who were diagnosed at the Department of Oral and Maxillofacial Surgery and 70 healthy individuals who at the Department of Medical Center of The First Affiliated Hospital of Zhengzhou University from August 2019 to January 2020 were recruited in The study. According to the random number table method, all subjects were divided into a test group (n=96) and a verification group (n=50). Subjects in the test group consisted of 51 OSCC patients and 45 healthy subjects and subjects in the verification group included 25 OSCC patients and 25 healthy individuals. Serum samples and clinical data of each of the subjects were collected. The serum samples were analyzed by ultra-high-performance liquid chromatography quadrupole-Orbitrap high resolution accurate mass spectrometry. Principal component analysis, orthogonal partial least square discrimination analysis and t-test were used to profile the differential metabolites in the test group. Pathway analysis of differential metabolites was performed. In addition, binary logistic regression analysis and receiver operating characteristic analysis were used in order to establish the potential diagnostic panel. Results: Twenty-one endogenous differential metabolites were identified showing significant association with OSCC. Results of pathway analysis suggested that OSCC associated with lipid metabolism and amino acid metabolism (P<0.05). A novel diagnostic panel consisting of lysophosphatidylcholine (LysoPC) (16∶0/0∶0), LysoPC[18∶1(9z)/0∶0], taurine and D-glutamic acid was defined. The panel performed a high area under the receiver operating characteristic curve (0.998, 95%CI: 0.994-0.999, P<0.05). Conclusions: There were obvious lipid and amino acid metabolism disorders in OSCC patients. It was an effective method to establish a diagnostic model by metabolomics.

Stromal DLK1 promotes proliferation and inhibits differentiation of the intestinal epithelium during development.

The stem/progenitor cells of the developing intestine are biologically distinct from their adult counterparts. Here, we examine the microenvironmental cues that regulate the embryonic stem/progenitor population, focusing on the role of Notch pathway factor delta-like protein-1 (DLK1). mRNA-seq analyses of intestinal mesenchymal cells (IMCs) collected from embryonic day 14.5 (E14.5) or adult IMCs and a novel coculture system with E14.5 intestinal epithelial organoids were used. Following addition of recombinant DLK1 (rDLK) or Dlk1 siRNA (siDlk1), epithelial characteristics were compared using imaging, replating efficiency assays, qPCR, and immunocytochemistry. The intestinal phenotypes of littermate Dlk1+/+ and Dlk1-/- mice were compared using immunohistochemistry. Using transcriptomic analyses, we identified morphogens derived from the embryonic mesenchyme that potentially regulate the developing epithelial cells, to focus on Notch family candidate DLK1. Immunohistochemistry indicated that DLK1 was expressed exclusively in the intestinal stroma at E14.5 at the top of emerging villi, decreased after birth, and shifted to the intestinal epithelium in adulthood. In coculture experiments, addition of rDLK1 to adult IMCs inhibited organoid differentiation, whereas Dlk1 knockdown in embryonic IMCs increased epithelial differentiation to secretory lineage cells. Dlk1-/- mice had restricted Ki67+ cells in the villi base and increased secretory lineage cells compared with Dlk1+/+ embryos. Mesenchyme-derived DLK1 plays an important role in the promotion of epithelial stem/precursor expansion and prevention of differentiation to secretory lineages in the developing intestine.NEW & NOTEWORTHY Using a novel coculture system, transcriptomics, and transgenic mice, we investigated differential molecular signaling between the intestinal epithelium and mesenchyme during development and in the adult. We show that the Notch pathway factor delta-like protein-1 (DLK1) is stromally produced during development and uncover a new role for DLK1 in the regulation of intestinal epithelial stem/precursor expansion and differentiation to secretory lineages.

The Balance of Stromal BMP Signaling Mediated by GREM1 and ISLR Drives Colorectal Carcinogenesis.

BACKGROUND & AIMS: Cancer-associated fibroblasts (CAFs), key constituents of the tumor microenvironment, either promote or restrain tumor growth. Attempts to therapeutically target CAFs have been hampered by our incomplete understanding of these functionally heterogeneous cells. Key growth factors in the intestinal epithelial niche, bone morphogenetic proteins (BMPs), also play a critical role in colorectal cancer (CRC) progression. However, the crucial proteins regulating stromal BMP balance and the potential application of BMP signaling to manage CRC remain largely unexplored. METHODS: Using human CRC RNA expression data, we identified CAF-specific factors involved in BMP signaling, then verified and characterized their expression in the CRC stroma by in situ hybridization. CRC tumoroids and a mouse model of CRC hepatic metastasis were used to test approaches to modify BMP signaling and treat CRC. RESULTS: We identified Grem1 and Islr as CAF-specific genes involved in BMP signaling. Functionally, GREM1 and ISLR acted to inhibit and promote BMP signaling, respectively. Grem1 and Islr marked distinct fibroblast subpopulations and were differentially regulated by transforming growth factor ß and FOXL1, providing an underlying mechanism to explain fibroblast biological dichotomy. In patients with CRC, high GREM1 and ISLR expression levels were associated with poor and favorable survival, respectively. A GREM1-neutralizing antibody or fibroblast Islr overexpression reduced CRC tumoroid growth and promoted Lgr5+ intestinal stem cell differentiation. Finally, adeno-associated virus 8 (AAV8)-mediated delivery of Islr to hepatocytes increased BMP signaling and improved survival in our mouse model of hepatic metastasis. CONCLUSIONS: Stromal BMP signaling predicts and modifies CRC progression and survival, and it can be therapeutically targeted by novel AAV-directed gene delivery to the liver.

[Analysis of the relationship between the changes of lung function and serum proinflammatory cytokines in workers occupationally exposed to toluene diisocyanate].

Objective: To analyze the correlation between the changes of lung function and serum proinflammatory cytokines in workers occupationally exposed to toluene diisocyanate (TDI), and to explore the evaluation index of respiratory toxicity of TDI. Methods: In October 2014, 61 male workers engaged in TDI synthesis process, purification process, packaging process and the above production process in a TDI factory in western China were selected as TDI exposure group; 62 male enterprise managers who were not exposed to TDI and other known allergenic chemicals were selected as control group, which were matched at the age of workers in exposure group. The questionnaire survey obtained information such as gender, length of service, age, occupational history, exposed length of service and so on. The lung function indexes [forced expiratory volume in the first second (FEV1), forced vital capacity (FVC), FEV1/FVC] and serum levels of interleukin (IL)-1 ß, IL-6, IL-8, tumor necrosis factor (TNF)-α, macrophage inflammatory factor-1 ß, monocyte chemoattractant factor-1 and vascular endothelial growth factor were measured. The urine was collected after the weekend shift, and the concentration of (TDA), the metabolite of TDI, was determined as the index of internal exposure. Spearman rank correlation was used to analyze the correlation between cytokines and lung function indexes, and multivariate linear regression was used to analyze the changes of lung function indexes and cytokines with TDI exposure concentration and time. Results: The median age (P5-P95) of the exposed group and the control group was 36.5 (24.0-51.0) and 38.0 (24.0-50.0) years, respectively. In the exposed group, the median length of service (P5-P95) was 6.94 (0.97-26.33) years, and the median concentration of TDA in urine was 15.56 (2.28-112.16) ng/ml. The three indexes of lung function, FVC, FEV1, FEV1/FVC and the levels of serum IL-8 and TNF-α were significantly lower than those in the control group (P<0.01). With the increase of exposure concentration and exposure time, the level of serum TNF-α, FVC and FEV1 decreased, and showed a good dose-effect and time-effect relationship (all Ptrend values< 0.05). Serum IL-8 and TNF-α were positively correlated with FVC, FEV1 and FEV1/FVC (all P values<0.01). Conclusion: The levels of serum inflammatory factors IL-8 and TNF-α in worker exposed to TDI are related to lung function indexes, which can be used as early evaluation indexes of respiratory toxicity induced by TDI.

Apatinib ameliorates doxorubicin-induced migration and cancer stemness of osteosarcoma cells by inhibiting Sox2 via STAT3 signalling.

BACKGROUND: Osteosarcoma, ranking as the second primary cause of cancer-related death, is the most common type of bone cancer. Doxorubicin (DOX) is used as a first-line treatment for osteosarcoma; however, the tumour recurrence rate remains high. Recent studies have suggested that DOX-induced migration and stemness in osteosarcoma cells might be the primary reason of recurrence and drug resistance. However, the underlying mechanisms remain unclear. Therefore, it is urgent to explore novel effective treatments to overcome DOX-induced drug resistance of osteosarcoma. METHODS: Osteosarcoma cells KHOS and U2OS were treated with DOX and apatinib (AP) alone or in combination. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and colony formation assays were performed to evaluate effects on proliferation. Flow cytometry analysis was carried out to assess cell apoptosis. Cell migration was determined by the transwell assay. Cancer stemness was detected by flow cytometry analysis of CD133+ cells and sphere-formation assay. Western blot assay was used to measure the expression of E-cadherin, N-cadherin, vimentin, matrix metalloproteinase 9 (MMP-9), signal transducer and activator of transcription 3 (STAT3)/p-STAT3, SRY-box2 (Sox2) and octamer-binding protein 4 (Oct4), and Nanog in treated osteosarcoma cells. RESULTS: Herein, we revealed that AP treatment significantly enhanced the sensitivity of osteosarcoma cells to DOX, reversed the DOX-induced stemness phenotype and metastasis capacity of osteosarcoma cells, and abolished the upregulation of p-STAT3, Sox2, Oct4, and Nanog. We further demonstrated that AP reversed DOX-induced stemness and migration of osteosarcoma cells through Sox2. CONCLUSION: These results suggested that AP significantly abolished the DOX-induced stemness phenotype and metastasis capacity in osteosarcoma cells by inhibiting Sox2 via STAT3 signalling. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: Our study indicates that Doxorubicin-based chemotherapeutics may simulate cancer stem cells (CSCs) properties in osteosarcoma, leading to the resistance of osteosarcoma. Apatinib can reduce the Doxorubicin-induced chemoresistance through STAT3/Sox2 pathway inactivation. This study represents that Apatinib may act as an effective chemotherapy sensitiser for reducing chemoresistance correlated with CSCs in osteosarcoma.