MiR-3074-5p suppresses non-small cell lung cancer progression by targeting the YWHAZ/Hsp27 axis.

Non-small cell lung cancer (NSCLC) accounts for more than 80% of lung cancer cases, and the 5-year survival rate of patients remains unsatisfactory. MicroRNAs (miRNAs) are small endogenous noncoding RNAs that are considered essential posttranscriptional regulators of tumorigenesis, including NSCLC. In this study, we aimed to investigate the biological role of miR-3074-5p in NSCLC cells and the underlying molecular mechanisms. We showed that miR-3074-5p expression was decreased in human NSCLC specimens and cell lines. Moreover, miR-3074-5p overexpression inhibited cell proliferation, migration and invasion and induced apoptosis and cell cycle arrest. In addition, miR-3074-5p overexpression not only suppressed tumor growth but also enhanced the antitumor effect of paclitaxel (PTX) on NSCLC cells in vitro and in vivo. A transcriptome sequencing assay revealed genes that were differentially expressed after miR-3074-5p overexpression, and among the genes whose expression levels were most significantly decreased, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) was a target of miR-3074-5p. The regulatory effect of miR-3074-5p on YWHAZ expression was verified by Western blotting and dual-luciferase reporter assays. The inhibition of A549 cell growth, migration and invasion was reversed by YWHAZ overexpression. Furthermore, we showed that PTX stimulated the expression of the YWHAZ and Hsp27 proteins and promoted the phosphorylation of Hsp27 (at S15 and S78). YWHAZ was confirmed to interact with Hsp27 in A549 cells, and downregulating YWHAZ expression promoted the degradation of the Hsp27 protein. Taken together, these results suggest that the miR-3074-5p/YWHAZ/Hsp27 axis may be a novel therapeutic target for NSCLC treatment.

Risk Factors Asscociated with Hypokalemia during Postanesthesia Recovery and Its Impact on Outcomes in Gynecological Patients: A Propensity Score Matching Study.

OBJECTIVE: This study aimed to explore the risk factors and outcomes of hypokalemia during the recovery period from anesthesia in the gynecological population. METHODS: This retrospective cohort study included 208 patients who underwent gynecological surgery at our institution between January 2021 and March 2022. Data were collected for each patient, including demographics, disease status, surgical data, and clinical information. Preoperative bowel preparation, postoperative gastrointestinal function, and electrolyte levels were compared between the two groups using propensity score matching (PSM). RESULTS: The incidence of hypokalemia (serum potassium level <3.5 mmol/L) during the recovery period from anesthesia was approximately 43.75%. After PSM, oral laxative use (96.4% vs. 82.4%, P=0.005), the number of general enemas (P=0.014), and the rate of ≥2 general enemas (92.9% vs. 77.8%, P=0.004) were identified as risk factors for hypokalemia, which was accompanied by decreased PaCO2 and hypocalcemia. There were no significant differences in postoperative gastrointestinal outcomes, such as the time to first flatus or feces, the I-FEED score (a scoring system was created to evaluate impaired postoperative gastrointestinal function), or postoperative recovery outcomes, between the hypokalemia group and the normal serum potassium group. CONCLUSION: Hypokalemia during postanesthesia recovery period occurred in 43.75% of gynecological patients, which resulted from preoperative mechanical bowel preparation; however, it did not directly affect clinical outcomes, including postoperative gastrointestinal function, postoperative complications, and length of hospital stay.

Non-sentinel lymph node metastases risk factors in patients with breast cancer with one or two sentinel lymph node macro-metastases.

Approximately 59 % of patients with breast cancer with one or two sentinel lymph nodes (1-2 SLN) macrometastases do not benefit from axillary lymph node dissection (ALND), which may also incur morbidities. It is necessary to evaluate the association between various clinicopathological characteristics and non-sentinel lymph node metastases (non-SLNM) in patients with breast cancer with 1-2 SLN macrometastases, and determine whether they 1-2 should avoid ALND. Eight electronic literature databases (PubMed, Embase, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Chinese Scientific Journal, Wanfang, and Chinese Biomedical Literature) were searched from their inception to June 30, 2023, and two reviewers independently extracted the data and assessed the risk of bias. Association strength was summarized using odds ratios (OR) and 95 % confidence intervals (CI). Heterogeneity was accounted for using a subgroup analysis. Publication bias was evaluated using funnel plots and Egger's test. There were 25 studies with 8021 participants, and 27 potential risk factors were evaluated. The risk factors for non-SLNM in patients with 1-2 SLN macrometastatic breast cancer include the following: factors of primary tumor: multifocality (OR (95 % CI (2.63 (1.96, 3.54))), tumor size ≥ T2 (2.64 (2.22, 3.14)), tumor localization (upper outer quad) (2.06 (1.23, 3.43)), histopathological grade (G3) (2.45 (1.70, 3.52)), vascular invasion (VI) (2.60 (1.35, 4.98)), lymphovascular invasion (LVI) (2.87 (1.80, 4.56)), perineural invasion (PNI) (3.16 (1.18,8.43)). Factors of lymph nodes: method of SLNs detected (blue dye) (3.85 (1.54, 9.60)), SLN metastasis ratio ≥0.5 (2.79 (2.24, 3.48)), two positive SLNs (3.55, (2.08, 6.07)), zero negative SLN (3.72 (CI 2.50, 4.29)), extranodal extension (ENE) (4.69 (2.16, 10.18)). Molecular typing: Her-2 positive (2.08 (1.26, 3.43)), Her-2 over-expressing subtype (1.83 (1.22, 2.73)). Factors of examination/inspection: axillary lymph nodes (ALNs) positive on imaging (3.18 (1.68, 6.00)), cancer antigen 15-3 (CA15-3) (4.01 (2.33,6.89)), carcinoembryonic antigen (CEA) (2.13 (1.32-3.43)). This review identified the risk factors for non-SLNM in patients with 1-2 SLN macrometastatic breast cancer. However, additional studies are needed to confirm the above findings owing to the limited number and types of studies included.

Deep vein thrombosis in a patient with Cronkhite-Canada syndrome: a complex case report.

BACKGROUND: Cronkhite-Canada syndrome (CCS) is a rare disease characterized by generalized gastrointestinal polyps, ectodermal abnormalities and variable gastrointestinal symptoms. Few cases to date have described complications with deep vein thrombosis (DVT). Here we reported a rare case of CCS concomitant with DVT. The patient's clinical details, endoscopic findings, safety, and efficacy are reported. CASE PRESENTATION: A 58-year-old patient was admitted to our hospital with recurrent diarrhea, overall abnormal appearance, including hyperpigmentation, hair loss and onychodystrophy, and multiple polyps distributed along the gastrointestinal tract except the esophagus. After considerable assessment, the patient was diagnosed with CCS. She was also diagnosed with concurrent DVT, nephrotic syndrome, and infectious enteritis during the course of disease. After treatment with a combination of methylprednisolone, mesalazine, antibiotics, rivaroxaban, and nutritional support during the 24 months of following the patient in this case, the clinical manifestations and endoscopic findings reached complete remission two years after the diagnosis. CONCLUSION: To our knowledge, this study is the first case of CCS complicated with DVT reported in China. Although rare, it is important to consider that DVT may occur after CCS and that it is vital to conduct careful follow-up.

Global burden of atrial fibrillation/atrial flutter and its attributable risk factors from 1990 to 2019.

AIMS: The aim of this study was to estimate the global burden of atrial fibrillation (AF)/atrial flutter (AFL) and its attributable risk factors from 1990 to 2019. METHODS AND RESULTS: The data on AF/AFL were retrieved from the Global Burden of Disease Study (GBD) 2019. Incidence, disability-adjusted life years (DALYs), and deaths were metrics used to measure AF/AFL burden. The population attributable fractions (PAFs) were used to calculate the percentage contributions of major potential risk factors to age-standardized AF/AFL death. The analysis was performed between 1990 and 2019. Globally, in 2019, there were 4.7 million [95% uncertainty interval (UI): 3.6 to 6.0] incident cases, 8.4 million (95% UI: 6.7 to 10.5) DALYs cases, and 0.32 million (95% UI: 0.27 to 0.36) deaths of AF/AFL. The burden of AF/AFL in 2019 and their temporal trends from 1990 to 2019 varied widely due to gender, Socio-Demographic Index (SDI) quintile, and geographical location. Among all potential risk factors, age-standardized AF/AFL death worldwide in 2019 were primarily attributable to high systolic blood pressure [34.0% (95% UI: 27.3 to 41.0)], followed by high body mass index [20.2% (95% UI: 11.2 to 31.2)], alcohol use [7.4% (95% UI: 5.8 to 9.0)], smoking [4.3% (95% UI: 2.9 to 5.9)], diet high in sodium [4.2% (95% UI: 0.8 to 10.5)], and lead exposure [2.3% (95% UI: 1.3 to 3.4)]. CONCLUSION: Our study showed that AF/AFL is still a major public health concern. Despite the advancements in the prevention and treatment of AF/AFL, especially in regions in the relatively SDI quintile, the burden of AF/AFL in regions in lower SDI quintile is increasing. Since AF/AFL is largely preventable and treatable, there is an urgent need to implement more cost-effective strategies and interventions to address modifiable risk factors, especially in regions with high or increased AF/AFL burden.

Different positive end expiratory pressure and tidal volume controls on lung protection and inflammatory factors during surgical anesthesia.

BACKGROUND: Mechanical ventilation can lead to the severe impairment of the metabolic pathway of alveolar surfactants, inactivating alveolar surfactants and significantly reducing lung-chest compliance. The cardiopulmonary function of elderly patients usually reduced to a certain extent, and there are lung complications after surgical anesthesia, just like lung barotrauma caused by mechanical ventilation, atelectasis and postoperative hypoxemia. AIM: To investigate the effects of different positive end expiratory pressures (PEEPs) and tidal volumes (VTs) on respiratory function, the degree of the inflammatory response and hemodynamic indexes in patients undergoing surgery under general anesthesia. METHODS: A total of 120 patients undergoing surgery for gastric or colon cancer under general anesthesia in Xinghua People's Hospital from January 2017 to January 2021 were randomly divided into Group A and Group B, with 60 cases in each group. The ventilation mode in Group A was VT (6.0 mL/kg) + PEEP (5.0 cmH2O), while that in Group B was VT (6.0 mL/kg) + PEEP (8.0 cmH2O). Blood gas parameters, respiratory mechanical parameters, inflammatory response indicators, hemodynamic indicators and related complications were compared between the two groups. RESULTS: There were no significant differences in PaCO2, PaO2, oxygen or the examined indexes at T0 between group A and group B (P > 0.05). The measured PaO2 value of patients in group A at T3 was higher than that in group B, and the difference was significant (P < 0.05). There were no significant differences in peak airway pressure (Ppeak), mean airway pressure or dynamic pulmonary compliance (Cdyn) at T0 between group A and group B (P > 0.05). The measured Ppeak value of patients in group A at T1 was higher than that in group B, and the difference was significant (P < 0.05). The measured Cdyn value at T1 and T2 was greater than that in group B (P < 0.05). Before surgery, there were no significant differences in tumor necrosis factor-α (TNF-α), interleukin (IL)-6 or IL-10 between group A and group B (P > 0.05). After 4 h, the measured values of TNF-α and IL-6 in group A were lower than those in group B, and the differences were significant (P < 0.05). The IL-10 Level in group A was higher than that in group B (P < 0.05). At T0, there were no significant differences in cardiac output, cardiac index (CI), stroke volume index (SVI) or mean arterial pressure between group A and group B (P > 0.05). The measured values of CI and SVI at T2 in patients in group A were higher than those in group B, and the differences were significant (P < 0.05). CONCLUSION: For patients undergoing surgery for gastric or colon cancer under general anesthesia, the VT (6.0 mL/kg) + PEEP (5.0 cmH2O) regimen was more effective than the VT (6.0 mL/kg) + PEEP (8.0 cmH2O) regimen in protecting the lung function and ventilatory function of patients, and it had better effects on maintaining hemodynamic stability and reducing inflammatory reactions.

Pseudomonas aeruginosa heteroresistance to levofloxacin caused by upregulated expression of essential genes for DNA replication and repair.

Pseudomonas aeruginosa (P. aeruginosa), a common cause of severe chronic infections, has developed heteroresistance to several antibiotics, thus hindering successful treatment. In this study, we aimed to investigate the characteristics and mechanisms underlying levofloxacin (LVX) heteroresistance in P. aeruginosa PAS71 and PAS81 clinical isolates using a combination of physiological and biochemical methods, bacterial genomics, transcriptomics, and qRT-PCR. The six P. aeruginosa strains, namely PAS71, PAS72, PAS81, PAS82, ATCC27853, and PAO1, were studied. The Kirby-Bauer (K-B), minimum inhibitory concentration (MIC) test, and population analysis profile (PAP) experimental results showed that PAS71, PAS81, ATCC27853, and PAO1 were heteroresistant to LVX, with MIC of 0.25, 1, 0.5, and 2 µg/ml, respectively; PAS72 and PAS82 were susceptible to LVX with a MIC of 0.25 and 0.5 µg/ml, respectively. The resistance of PAS71 and PAS81 heteroresistant subpopulations was unstable and had a growth fitness cost. Genomic and transcriptomic results proved that the unstable heteroresistance of PAS71 and PAS81 was caused by elevated expression of essential genes involved in DNA replication and repair, and homologous recombination, rather than their genomic single-nucleotide polymorphism (SNP) and insertion-deletion (InDel) mutations. Additionally, PAS71 and PAS81 enhanced virulence and physiological metabolism, including bacterial secretion systems and biosynthesis of siderophore group nonribosomal peptides, in response to LVX stress. Our results suggest that the upregulation of key genes involved in DNA replication and repair, and homologous recombination causes unstable heteroresistance in P. aeruginosa against LVX. This finding provides novel insights into the occurrence and molecular regulation pathway of P. aeruginosa heteroresistant strains.

Responsive nanoplatform for persistent luminescence "turn-on" imaging and "on-demand" synergistic therapy of bacterial infection.

Multifunctional nanotheranostic platforms are emerging for the treatment of bacterial infections. Uncontrollable drug release and poor response in target location leads to inefficient therapy and failure to offer timely antibacterial monitoring. Here, we report a multifunctional nanoplatform that can be triggered by the bacterial microenvironment for effective bacterial killing and high-sensitive persistent luminescence (PL) "turn-on" imaging. Hyaluronic acid (HA) is grafted on the surface of mesoporous silica-coated persistent luminescence nanoparticles (PLNPs@MSN) loaded with cinnamaldehyde (CA). Further in situ growth of MnO2 shells gives PLNPs@MSN@CA-HA-MnO2 (PMC-HA-MnO2). MnO2 shell of PMC-HA-MnO2 can be reduced to Mn2+ by the H2O2 in the bacterial microenvironment to trigger persistent luminescence (PL) "turn-on" imaging along with chemodynamic therapy (CDT). Meanwhile, HA can response to bacterially secreted hyaluronidase to make the packaged CA release controllable and "on-demand". Consequently, PMC-HA-MnO2 enables effective response to bacterial-infected region, ensuring high-sensitive "turn-on" imaging, synergistic CDT, accurate targeting and "on-demand" CA release to give great antibacterial effect. This nanoplatform has great potential for the diagnosis and treatment of multidrug-resistant bacterial infection with high specificity and efficiency.

[Probabilistic Risk Assessment of Arsenic Exposure Through Drinking Water Intake in Chinese Residents].

The total arsenic concentrations of the three main drinking water types in China were determined through a systematic literature review. The distribution models of drinking water exposure parameters for different age groups were obtained using the regression method. The carcinogenic and non-carcinogenic risks of different population groups caused by arsenic exposure through different drinking water types were evaluated by a probabilistic risk assessment. The results showed that the geometric mean of total arsenic content in all the drinking water samples in China was (13.0±38.1) µg·L-1. The highest arsenic content was found in the Inner Mongolia Autonomous Region, followed by Guangxi and Shanxi. Based on the relevant standards for drinking water quality, the probability exceeding the threshold value of groundwater and source water is 23.2% and 18.4%, respectively. According to the results of non-carcinogenic risk assessment, the probability of the residents drinking well water and surface water from the water source area exceeding the daily average exposure dose threshold was 24.0% and 19.5%, respectively. According to the carcinogenic risk assessment, the median of carcinogenic risk caused by arsenic in drinking water in China was 3.22×10-5, which is acceptable. The population group of 18-45 years old had the highest risk, and the median LCR was 1.37×10-5. There was still a certain probability that the LCR of drinking well water and surface water would exceed the acceptable risk level recommended by the US EPA. In conclusion, the potential health risks caused by arsenic exposure through drinking water intake exist among Chinese residents. Further control of the arsenic concentration in drinking water is required to reduce the health risk and improve the safety of drinking water. Meanwhile, it is suggested to strengthen the research on risk threshold to provide a scientific basis for the residents' health protection.

Multisystem involvement Langerhans cell histiocytosis in an adult: A case report.

BACKGROUND: Langerhans cell histiocytosis (LCH) is a rare condition wherein Langerhans cells proliferate abnormally, adversely impacting organs including lymph nodes, bones, skin, lungs, and pituitary gland. The LCH disease course varies widely among patients from a self-limiting condition to one that progresses rapidly and culminates in death. It is uncommon for multisystem LCH to be observed in adults. Herein we describe a woman suffering from multi-system LCH involvement. CASE SUMMARY: A 37-year old Chinese woman was admitted to the hospital in June 2019 suffering from dyspnea that had progressed over the course of 5 years. Her medical history included: central diabetes insipidus (DI) that had been treated via radiotherapy, desmopressin acetate, and bromocriptine; bilateral pneumothorax with two surgeries having been performed to remove bullae; and autoimmune hepatitis that had been unsuccessfully treated using a combination of methylprednisolone and mycophenolate mofetil. A chest computed tomography (CT) scan revealed the presence of multiple pulmonary cysts of varying sizes. We re-analyzed right pulmonary bullae samples that had been removed in 2014, performed a systematic 18F-FDG PET/CT analysis, and convened a multidisciplinary medical team to diagnose and treat this patient. As a result, we were able to eventually diagnose this patient with LCH that was not associated with BRAF-V600E mutations. CONCLUSION: We hope to emphasize the importance of systemic evaluation and of cooperation between multidisciplinary physicians with the goal of improving awareness and detection of this orphan disease.

Expression of TAZ, YAP, and ß-catenin in cervical squamous cell carcinoma and their clinical significance.

YAP/TAZ and ß-catenin are important effectors in the Hippo and Wnt signaling pathways, respectively, which are involved in the development of human tumors. Using immunohistochemistry, the expression levels of the three proteins were determined in 151 cervical tissue samples (including 28 normal cervical, 31 cervical intraepithelial neoplasia, and 92 cervical squamous cell carcinoma [CSC] tissues), which were excised or biopsied by surgery. The results showed that the three proteins were differently expressed in normal, precancerous, and CSC tissues, and ß-catenin expression positively correlated with both YAP and TAZ expression. By analyzing the relationships between YAP, TAZ, and ß-catenin expression and the clinicopathologic characteristics of patients with CSC, we found that YAP was related to the depth of invasion > 1/2, the diameter of the tumor > 4 cm, and positive lymph nodes; while TAZ and ß-catenin were related to the depth of invasion > 1/2 and positive lymph nodes. Regarding the prognostic factors of patients with CSC, Kaplan-Meier univariate and Cox multivariate regression analysis showed that there were significant correlations between lymph node infiltration; expression of YAP, TAZ, and ß-catenin; and patient mortality (P < 0.05), all of which were independent factors influencing mortality (OR > 1).

Gut microbiota contributes to the distinction between two traditional Chinese medicine syndromes of ulcerative colitis.

BACKGROUND: Ulcerative colitis (UC) is considered to be closely associated with alteration of intestinal microorganisms. According to the traditional Chinese medicine (TCM) theory, UC can be divided into two disease syndromes called Pi-Xu-Shi-Yun (PXSY) and Da-Chang-Shi-Re (DCSR). The relationships among gut microbiota, TCM syndromes, and UC pathogenesis have not been well investigated. AIM: To investigate the role of gut microbiota in UC and the distinction of microbiota dysbiosis between PXSY and DCSR syndromes. METHODS: From May 2015 to February 2016, UC patients presenting to LongHua Hospital who met the established inclusion and exclusion criteria were enrolled in this retrospective study. Fresh stool specimens of UC patients with PXSY or DCSR were collected. The feces of the control group came from the health examination population of Longhua Hospital. The composition of gut bacterial communities in stool samples was determined by the pyrosequencing of 16S ribosomal RNA. The high-throughput sequencing reads were processed with QIIME, and biological functions were predicted using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States. RESULTS: The composition of gut bacterial communities in 93 stool samples (30 healthy controls, 32 patients with PXSY syndrome, and 31 patients with DCSR syndrome) was determined by the pyrosequencing of 16S ribosomal RNA. Beta diversity showed that the composition of the microbiota was different among the three groups. At the family level, Porphyromonadaceae, Rikeneliaceae, and Lachnospiraceae significantly decreased while Enterococcus, Streptococcus, and other potential pathogens significantly increased in UC patients compared to healthy subjects. At the genus level, Parabacteroides, Dorea, and Ruminococcus decreased while Faeca-libacterium showed increased abundance in UC compared to healthy controls. Five differential taxa were identified between PXSY and DCSR syndromes. At the genus level, a significantly increased abundance of Streptococcus was observed in DCSR patients, while Lachnoclostridium increased in PXSY patients. The differential functional pathways of the gut microbiome between the PXSY and DCSR groups mainly included lipid metabolism, immunity, and the metabolism of polypeptides. CONCLUSION: Our study suggests that the gut microbiota contributes to the distinction between the two TCM syndromes of UC.

Potential involvement of heat shock proteins in pancreatic-duodenal homeobox-1-mediated effects on the genesis of gastric cancer: A 2D gel-based proteomic study.

AIM: To identify functional proteins involved in pancreatic-duodenal homeobox-1 (PDX1)-mediated effects on gastric carcinogenesis. METHODS: A PDX1-overexpressed model was established by transfecting gastric cancer cell line SGC7901 with pcDNA3.1(+)-PDX1 vector (SGC-PDX1). Transfection with empty pcDNA3.1 vector (SGC-pcDNA) served as control. Comparative protein profiles of the two groups were analyzed by two-dimensional electrophoresis based-proteomics (2DE gel-based proteomics). The differential proteins identified by 2DE were further validated by qRT-PCR and immunoblotting. Finally, co-immunoprecipitation was used to determine any direct interactions between PDX1 and the differential proteins. RESULTS: 2DE gel proteomics identified seven differential proteins in SGC-PDX1 when compared with those in SGC-pcDNA. These included four heat shock proteins (HSPs; HSP70p1B, HSP70p8, HSP60, HSP27) and three other proteins (ER60, laminin receptor 1, similar to epsilon isoform of 14-3-3 protein). Immunoblotting validated the expression of the HSPs (HSP70, HSP60, HSP27). Furthermore, their expressions were lowered to 80%, 20% and 24%, respectively, in SGC-PDX1, while PDX1 exhibited a 9-fold increase, compared to SGC-pcDNA. However, qRT-PCR analysis revealed that mRNA levels of the HSPs were increased in SGC-PDX1, suggesting that the expression of the HSPs was post-translationally regulated by the PDX1 protein. Finally, co-immunoprecipitation failed to identify any direct interaction between PDX1 and HSP70 proteins. CONCLUSION: This study demonstrates the potential involvement of HSPs in PDX1-mediated effects on the genesis of gastric cancer.

Solitary pleural myeloma diagnosed by semi-rigid thoracoscopy: A case report and literature review.

Multiple myeloma (MM) is characterized by abnormal proliferation of neoplastic plasma cells. Pleural effusion as an initial presentation of this disease is rare, as is true pleural myeloma. We herein present a case of solitary pleural myelomatous lesion in a 70-year-old male patient diagnosed by pleural biopsy via semi-rigid thoracoscopy followed by histopathological examination. Furthermore, a review of the related English literature identified 22 cases of pleural myeloma, only 3 of which were diagnosed by video-assisted thoracoscopy. To the best of our knowledge, this is the first reported case of a solitary pleural myelomatous lesion diagnosed by pleural biopsy via semi-rigid thoracoscopy. Patients with MM with pleural involvement, including the present case, appear to have a short survival despite aggressive treatment. Our patient received chemotherapy with bortezomib, epiadriamycin and dexamethasone; however, he deteriorated rapidly after one cycle of chemotherapy and succumbed to the disease 8 weeks after the initial presentation. According to our experience, semi-rigid thoracoscopy is an effective and safe method for obtaining a pleural specimen for histopathological evaluation.

New histopathologic and ultrastructural findings in Reis-Bücklers corneal dystrophy caused by the Arg124Leu mutation of TGFBI gene.

BACKGROUND: Reis-Bücklers corneal dystrophy (RBCD) was consistently reported as a corneal dystrophy only affected Bowman's layer and superficial corneal stroma, and superficial keratectomy was a recommendation surgery for treatment in literatures. The study reported new histopathological and ultrastructural findings in RBCD caused by the Arg124Leu mutation of transforming growth factor induced (TGFBI) gene in a four-generation Chinese pedigree. METHODS: Subjects including eight patients and seven unaffected family members received slit-lamp biomicroscopy and photography. DNA was obtained from all subjects, and exons 4 and 11 to 14 of TGFBI gene were analyzed by polymerase chain reaction and the products were sequenced. Anterior segment optical coherence tomography (AS OCT) and in vivo confocal microscopy were conducted for ten eyes of five patients. Based on the results of AS OCT and in vivo confocal microscopy, deep anterior lamellar keratoplasty (DLKP) using cryopreserved donor cornea was applied for four eyes of four patients. Four lamellar dystrophic corneal buttons were studied by light and transmission electron microscopy, and TGFBI immunohistochemistry. RESULTS: Eight patients had typical clinical manifestations of RBCD presenting recurrent painful corneal erosion starting in their early first decades, along with age-dependent progressive geographic corneal opacities. TGFBI sequencing revealed a heterozygous mutation, Arg124Leu in all eight patients. Anterior segment optical coherence tomography and in vivo confocal microscopy showed the dystrophic deposits involved not only in subepithelial and superficial stroma, but also in mid- or posterior stroma in four examined advanced eyes. Light microscopy showed Bowman's layer was absent, replaced by abnormal deposits stain bright red with Masson's trichrome. In superficial cornea, the deposits stacked and produced three to five continuous bands parallel to the corneal collagen lamellae. In mid- to posterior stroma, numerous granular or dot- like aggregates were heavily scattered, and most of them presented around the nuclei of stromal keratocytes. Transmission electron microscopy revealed the multiple electron-dense rod-shaped deposits aggregated and formed a characteristic pattern of three to five continuous bands in superficial cornea, which were similar to those seen under light microscopy. In mid- to posterior stroma, clusters of rod-shaped bodies were scattered extracellular or intracellular of the stromal keratocytes between the stromal lamellae suggesting the close relationship between mutated proteins and keratocyte. CONCLUSIONS: The study offer evidences indicating DLKP is a viable treatment option for advanced RBCD to avoid recurrence, and the mutated TGFBIp in dystrophic corneas are of keratocytes origin.

Control of Trx1 redox state modulates protection against methyl methanesulfonate-induced DNA damage via stabilization of p21.

Thioredoxin 1 (Trx1) is known to play an important role in protecting against cell death. However, the mechanism for control of Trx1 in cell death resulting from DNA damage has not been fully investigated. In this study, we used the DNA-damaging agent methyl methanesulfonate (MMS) to investigate the protective effects of Trx1 against DNA damage and cell death in HEK293 cells. We found that MMS application caused dose-dependent changes in the Trx1 redox state determined by redox western blotting. At lower concentrations, both reduced and oxidized Trx1 were observed, whereas the reduced band was fully oxidized at the higher concentration. Trx1 overexpression and small interfering RNA knockdown in cells revealed that reduced Trx1 after exposure to lower doses of MMS attenuated DNA damage, assessed by comet assay, and level of the DNA-damage marker histone γ-H2AX, possibly through scavenging intracellular ROS and an increase in p21 protein level via enhancing its stability. However, oxidized Trx1 lost its protective ability to DNA damage in response to higher concentration of MMS. Corresponding to the redox state control of Trx1, cell death induced by different dose of MMS was also found, by inhibiting phosphorylations of p38 and 4E-BP1. These results indicate that reduced Trx1 plays important protective roles against MMS-induced DNA damage and cell death, suggesting that cell protection is regulated by the intracellular redox state. Control of the redox state of Trx1 and its regulating proteins may offer a novel therapeutic strategy for the control of cancer.

Autophagy is essential for ultrafine particle-induced inflammation and mucus hyperproduction in airway epithelium.

Environmental ultrafine particulate matter (PM) is capable of inducing airway injury, while the detailed molecular mechanisms remain largely unclear. Here, we demonstrate pivotal roles of autophagy in regulation of inflammation and mucus hyperproduction induced by PM containing environmentally persistent free radicals in human bronchial epithelial (HBE) cells and in mouse airways. PM was endocytosed by HBE cells and simultaneously triggered autophagosomes, which then engulfed the invading particles to form amphisomes and subsequent autolysosomes. Genetic blockage of autophagy markedly reduced PM-induced expression of inflammatory cytokines, e.g. IL8 and IL6, and MUC5AC in HBE cells. Mice with impaired autophagy due to knockdown of autophagy-related gene Becn1 or Lc3b displayed significantly reduced airway inflammation and mucus hyperproduction in response to PM exposure in vivo. Interference of the autophagic flux by lysosomal inhibition resulted in accumulated autophagosomes/amphisomes, and intriguingly, this process significantly aggravated the IL8 production through NFKB1, and markedly attenuated MUC5AC expression via activator protein 1. These data indicate that autophagy is required for PM-induced airway epithelial injury, and that inhibition of autophagy exerts therapeutic benefits for PM-induced airway inflammation and mucus hyperproduction, although they are differentially orchestrated by the autophagic flux.

Intravenous injection of l-BMAA induces a rat model with comprehensive characteristics of amyotrophic lateral sclerosis/Parkinson-dementia complex.

Non-protein amino acid beta-N-methylamino-l-alanine (l-BMAA) is a neurotoxin that was associated with the high incidence of Amyotrophic Lateral Sclerosis/Parkinson-Dementia Complex (ALS/PDC) in Guam. This neurotoxin has been implicated as a potential environmental factor in amyotrophic lateral sclerosis, Alzheimer's disease and other neurodegenerative diseases, and was found to accumulate in brain tissues of ALS/PDC patients. It is extremely important to establish a reliable animal model that has the comprehensive characteristics of ALS/PDC for studying mechanisms underlying neurodegeneration, and exploring effective therapies. However, very few good animal models that mimic ALS/PDC have been established. In this study, an ideal rat model that mimicked most characteristics of ALS/PDC was established by administering continuous intravenous (i.v.) injections of neurotoxic l-BMAA. Based on the data obtained, it was demonstrated that continuous i.v. injections of l-BMAA induced mitochondrial morphology and structural changes, astrogliosis, motor neuronal death, and other relative functional changes, which led to the overexpression of pro-inflammatory cytokines cyclooxygenase-2 (COX-2), nuclear factor kappa B (NF-κB) and tumor necrosis factor-alpha (TNF-α), and resulted in the upregulation of glycogen synthase kinase-3 (GSK3), downregulation of astrocytic glutamate transporter-1 (GLT-1), accumulation of microtubule-associated protein tau and cytosolic aggregates of TAR DNA-binding protein-43 (TDP-43) in degenerating motor neurons. These results suggest that this model could be used as a useful tool for the mechanistic and therapeutic study of ALS/PDC.

Transplantation of bone marrow-derived mesenchymal stem cells facilitates epithelial repair and relieves the impairment of gastrointestinal function in a rat model of enteritis.

BACKGROUND: To examine whether the bone marrow-derived MSCs (BM-MSCs) could facilitate epithelial repair and thereby reduce impairment of gastrointestinal structure and function in chronic murine enteritis induced by indomethacin (IDM). METHODS: MSCs were isolated from young Sprague-Dawley rats. After in vitro expansion and characterization, BM-MSCs were labelled with the fluorescent dye PKH26 and transfused, via the tail veins, into rats with enteritis induced by IDM. The controls were infused with sterile saline. The homing and differentiation of the transplanted BM-MSCs were tracked by means of fluorescent staining. The clinical symptoms of the IDM-treated rats were assessed, and the macroscopic and microscopic histological evaluations of the intestines were performed. RESULTS: Compared to controls that received saline infusion, BM-MSCs treated rats showed lower scores of weight loss, stool consistency, and stool blood. The PKH26-labelled cells resided at the injured intestine, where they co-localize with the proliferating cell nuclear antigen (PCNA), Lgr-5, and Msi-1. The BM-MSCs treated rats showed significantly higher intestinal villi with larger areas relative to the saline-treated rats. CONCLUSION: The transplanted BM-MSCs are able to recognize the injured intestine, where they proliferate and transdifferentiate into intestinal stem cells which repair the injured intestinal tissues. Therefore, BM-MSCs are able to relieve the impairment of gastrointestinal function in IMD-treated rats.