Trigonelline hydrochloride attenuates silica-induced pulmonary fibrosis by orchestrating fibroblast to myofibroblast differentiation.

BACKGROUND: Silicosis represents a paramount occupational health hazard globally, with its incidence, morbidity, and mortality on an upward trajectory, posing substantial clinical dilemmas due to limited effective treatment options available. Trigonelline (Trig), a plant alkaloid extracted mainly from coffee and fenugreek, have diverse biological properties such as protecting dermal fibroblasts against ultraviolet radiation and has the potential to inhibit collagen synthesis. However, it's unclear whether Trig inhibits fibroblast activation to attenuate silicosis-induced pulmonary fibrosis is unclear. METHODS: To evaluate the therapeutic efficacy of Trig in the context of silicosis-related pulmonary fibrosis, a mouse model of silicosis was utilized. The investigation seeks to elucidated Trig's impact on the progression of silica-induced pulmonary fibrosis by evaluating protein expression, mRNA levels and employing Hematoxylin and Eosin (H&E), Masson's trichrome, and Sirius Red staining. Subsequently, we explored the mechanism underlying of its functions. RESULTS: In vivo experiment, Trig has been demonstrated the significant efficacy in mitigating SiO2-induced silicosis and BLM-induced pulmonary fibrosis, as evidenced by improved histochemical staining and reduced fibrotic marker expressions. Additionally, we showed that the differentiation of fibroblast to myofibroblast was imped in Trig + SiO2 group. In terms of mechanism, we obtained in vitro evidence that Trig inhibited fibroblast-to-myofibroblast differentiation by repressing TGF-ß/Smad signaling according to the in vitro evidence. Notably, our finding indicated that Trig seemed to be safe in mice and fibroblasts. CONCLUSION: In summary, Trig attenuated the severity of silicosis-related pulmonary fibrosis by alleviating the differentiation of myofibroblasts, indicating the development of novel therapeutic approaches for silicosis fibrosis.

Repeat resection versus percutaneous ablation for recurrent hepatocellular carcinoma: a meta-analysis.

Introduction: Both repeat resection (RR) and percutaneous ablation (PA) have been used for treating recurrent hepatocellular carcinoma (rHCC). Each method has its advantages and disadvantages. Aim: To compare the safety and effectiveness between RR and PA in patients with rHCC. Material and methods: Relevant articles published in the PubMed, Embase, Wanfang, and China National Knowledge Infrastructure (CNKI) databases published as of April 2022 were identified. Primary endpoints for this meta-analysis included patient overall survival (OS) and disease-free survival (DFS), whereas secondary endpoints included rates of repeat recurrence, complications, and the duration of hospitalization. Results: This meta-analysis included a total of 6 relevant studies. Pooled repeat recurrence rates were comparable between the PA and RR groups (p = 0.09), although the pooled 5-year DFS rate (p = 0.01), DFS duration (p = 0.02), and 3-year OS rate (p = 0.04) in the RR group were considerably higher than in the PA group. Pooled rates of both Grade 1/2 (p = 0.04) and Grade 3/4 (p = 0.001) complications, however, were significantly lower for patients who underwent PA as compared to patients who underwent RR. PA was associated with a significantly shorter hospitalization duration relative to RR in this patient cohort (p = 0.0002). Conclusions: According to the obtained findings, RR may be associated with better long-term disease control in rHCC patients than PA, whereas PA is associated with a better safety profile and a shorter duration of hospitalization.

Stenting with high-intensity focused ultrasound ablation for distal biliary obstruction caused by pancreatic carcinoma: a meta-analysis.

Introduction: Between 42% and 77% of patients with distal malignant biliary obstruction (MBO) suffer from pancreatic carcinoma (PC). Aim: To analyse the clinical efficacy of stenting accompanied by high-intensity focused ultrasound (HIFU) ablation in patients with distal MBO from PC. Material and methods: Relevant articles published through March 2021 were identified in the Pubmed, Cochrane Library, Embase, Wanfang, VIP, and CNKI databases. RevMan v5.3 and Stata v12.0 were used for the meta-analysis. Results: Twenty-nine articles were initially identified, and 5 of these were eventually included. These articles described 142 patients who underwent biliary stenting alone and 132 patients who underwent biliary stenting with HIFU ablation. The pooled Δ total bilirubin (TBIL) values were comparable between the 2 treatment groups (p = 0.10). The pooled stent dysfunction rate was significantly greater in the group with stenting alone (p = 0.03), and the pooled HR for the stent patency duration indicated that the duration of stent patency was increased in the stenting with HIFU ablation group (p < 0.0001). Overall survival rates were significantly longer in the stenting with HIFU ablation group (p < 0.0001). HIFU ablation was associated with an 80% pooled clinical response rate. The pooled cholangitis (p = 0.47) and pancreatitis (p = 0.56) rates were comparable between the 2 groups. Funnel plots did not reveal any significant evidence of endpoint-associated publication bias. Conclusions: Stenting with HIFU ablation increased both stent patency and overall survival in patients with distal MBO caused by PC compared to stenting alone.

Correlation between promoter methylation of the LDH-C4 gene and DNMT expression in breast cancer and their prognostic significance.

DNA methylation plays an important role in tumorigenesis and development. The potential of aberrant DNA methylation to act as a biomarker for tumor diagnosis and prognostic evaluation is currently being explored. Lactate dehydrogenase C4 (LDH-C4) is a member of the cancer/testis antigen family that is expressed in a broad range of human tumor types, with particularly high expression patterns observed in lung cancer, melanoma and breast cancer. However, whether the methylation of the promoter region of LDH-C4 can be used as a tumor marker and its association with prognosis remain poorly understood. The present study aimed to determine the potential of the methylation status of LDH-C4 and DNA methyltransferase (DNMT) expression to be biological markers for the prognostic evaluation of breast cancer. Methylation-specific PCR was conducted to evaluate alterations in the methylation levels of LDH-C4. Immunohistochemical analysis was performed to assess the expression levels of DNMTs in breast cancer tissues. The association between the methylation status of LDH-C4 or the expression of DNMTs, and clinical pathological parameters was also evaluated. The results of the present study revealed that the demethylation rate of LDH-C4 in breast cancer tissues was significantly increased compared with that of adjacent tissues, and associations were identified between the demethylation of LDH-C4 and the histological grade, estrogen receptor, progesterone receptor and HER-2 status, and lymph node metastasis. The level of LDH-C4 demethylation was negatively correlated with the expression of DNMTs. Demethylation of the LDH-C4 promoter and DNMT expression predicted an unfavorable prognosis of patients with breast cancer. In addition, demethylation of the LDH-C4 promoter, high expression of DNMT3a and DNMT3b, histological grade and lymph node metastasis were all discovered to be independent prognostic factors in patients with breast cancer. In conclusion, results of the present study indicated that the demethylation of LDH-C4 and DNMT expression levels may be closely associated with the occurrence and development of breast cancer, in addition to lymph node metastasis. Thus, both may be used to assist the clinical evaluation of prognosis.

Transarterial embolization with hepatectomy for ruptured hepatocellular carcinoma: a meta-analysis.

PURPOSE: To compare the clinical effectiveness between transarterial embolization (TAE) with staged hepatectomy (SH) and emergency hepatectomy (EH) for ruptured hepatocellular carcinoma (HCC). MATERIAL AND METHODS: Pubmed, Embase, and Cochrane Library databases were screened for eligible publications from the inception of the databases till February 2021. RESULTS: This meta-analysis included seven studies comprising 162 patients who underwent TAE with SH and 266 patients who underwent EH. The pooled intraoperative blood loss was less in the TAE with SH cohort, as compared to the EH cohort without significant difference (p = .20). The pooled blood transfer rate (p<.00001), blood transfer volume (p = .002), and 30-day patient death (p = .04) were all markedly reduced in the TAE with SH cohort versus the EH cohort. No significant differences in surgery duration (p = .27), hospital stay period (p = .81), complication rate (p = 0.92), disease-free survival (DFS) (p = .79), and overall survival (OS) (p = 0.28) were found between the two groups. CONCLUSIONS: Compared with EH for ruptured HCC, TAE with SH could effectively decrease intraoperative blood loss and 30-day mortality. However, the long-term DFS and OS might not be beneficial to preoperative TAE.

Stent Insertion With High Intensity-Focused Ultrasound Ablation for Biliary Obstruction Caused by Pancreatic Carcinoma: A Randomized Controlled Trial.

PURPOSE: This study was designed to assess the clinical efficacy of stent insertion with high intensity-focused ultrasound ablation (HIFUA) in patients with malignant biliary obstruction (MBO) as a consequence of pancreatic carcinoma (PC). MATERIALS AND METHODS: This was a single-center, open-label, prospective, randomized controlled trial. Consecutive patients with MBO caused by PC were randomly assigned to undergo stent insertion with or without HIFUA from June 2019 to February 2020. This study was registered at ClinicalTrials.gov (NCT03962478). RESULTS: In total, 92 patients were enrolled in this study and assigned to the stent-only (n=46) or combined (stent+HIFUA; n=46) treatment groups. Stent insertion was associated with a 100% technical success rate. For patients in the combination treatment group, 26, 18, and 2 patients underwent 2, 3, and 4 cycles of HIFUA, respectively. A positive clinical response to HIFUA treatment was noted in 38 patients (82.6%). Stent dysfunction was detected in 9 and 15 patients in the combination and stent-only groups, respectively (P=0.154), while median stent patency in these 2 groups was 188 and 120 days, respectively (P<0.001). All patients died over the course of the follow-up, with median survival periods of 218 and 140 days in the combination and stent-only treatment groups, respectively (P=0.001). The only detected predictor of prolonged survival was HIFUA treatment (P=0.004), and there were no significant differences in complication rates between these 2 treatment groups. CONCLUSION: A combination of stent insertion and HIFUA can improve stent patency and overall survival in patients suffering from MBO because of PC relative to stent insertion alone.

Repurposed Tocilizumab in Patients with Severe COVID-19.

The coronavirus disease 2019 (COVID-19) has caused a global pandemic, resulting in considerable morbidity and mortality. Tocilizumab, an inhibitor of IL-6, has been widely repurposed as a treatment of severely ill patients without robust evidence supporting its use. In this study, we aimed to systematically describe the effectiveness of treatment and prevention of the cytokine storms in COVID-19 patients with tocilizumab. In this multicentered retrospective and observational cohort study, 65 patients with COVID-19 receiving tocilizumab and 130 not receiving tocilizumab were propensity score matched at a ratio of 2:1 based on age, sex, and comorbidities from January 20, 2020 to March 18, 2020 in Wuhan, China. After adjusting for confounding, the detected risk for in-hospital death was lower in the tocilizumab group versus nontocilizumab group (hazard ratio = 0.47; 95% confidence interval = 0.25-0.90; p = 0.023). Moreover, use of tocilizumab was associated with a lower risk of acute respiratory distress syndrome (odds ratio = 0.23; 95% confidence interval = 0.11-0.45; p < 0.0001). Furthermore, patients had heightened inflammation and more dysregulated immune cells before treatment, which might aggravate disease progression. After tocilizumab administration, abnormally elevated IL-6, C-reactive protein, fibrinogen, and activated partial thromboplastin time decreased. Tocilizumab may be of value in prolonging survival in patients with severe COVID-19, which provided a novel strategy for COVID-19-induced cytokine release syndrome. Our findings could inform bedside decisions until data from randomized, controlled clinical trials become available.

Dexamethasone prevents the Epstein-Barr virus induced epithelial-mesenchymal transition in A549 cells.

Clinical data have shown that pulmonary interstitial fibrosis is likely to occur in the later stages of viral pneumonia. While viral infections are thought to cause chronic pulmonary interstitial inflammation and pulmonary fibrosis, it remains unclear if they promote pulmonary fibrosis by epithelial-mesenchymal transition (EMT). In this study, human epithelial cell line A549 has been used to model the infection of the Epstein-Barr virus (EBV) and the respiratory syncytial virus (RSV). Their differences were compared and the possible infection mechanisms analyzed by randomly assigning cells to one of five treatments. Exposure of the LMP1 is thought to be the key gene during EBV-induced EMT in the A549 cells. Enzyme-linked immunosorbent assay analysis revealed that the EBV infection was associated with the induction of a number of cytokines (interleukin-8 [IL-8], IL-13, tumor necrosis factor-α, and transforming growth factor-ß) and dexamethasone (DXM) could significantly prevent the phenotypic changes, and partly the mechanisms related with the IL-13 pathway. Surprisingly, different results were seen with the RSV infection as the A549 cells still displayed an epithelial morphology but the levels of E-cadherin, α-SMA, vimentin, and fibronectin did not change. This is the first study demonstrating the different reactions induced by different viruses, and the protective effects of DXM on the EBV-induced EMT in the A549 cells by partially inhibiting the IL-13 pathway. These findings suggest a novel mechanism, by which DXM or anti-IL-13 may delay the progression of pulmonary fibrosis by preventing the progress of EBV-induced EMT.

Influence of polymorphisms in the Wnt/ß-catenin pathway genes on hepatocellular carcinoma risk in a Chinese Han population.

The Wnt/ß-catenin pathway plays a vital role in initiating and sustaining hepatocellular carcinoma (HCC). However, few studies have investigated polymorphisms in the Wnt/ß-catenin signaling pathway genes in the Chinese Han population. The aim of the present retrospective study was to investigate the correlations between polymorphisms of the Wnt/ß-catenin signaling pathway genes (CTNNB1 and WNT2) and HCC susceptibility, development, and progression.Twenty tagging single nucleotide polymorphisms were chosen from HapMap data and genotyped in 320 patients with HCC, 320 chronic hepatitis B virus (HBV)-infected patients without HCC (N-HCC, including 95 liver cirrhosis, 164 chronic hepatitis B, and 61 asymptomatic HBV carriers), and 320 healthy controls. Associations between polymorphisms in the 2 Wnt/ß-catenin signaling pathway genes (CTNNB1 and WNT2) and HCC susceptibility, development, and progression were investigated.Genotype AA (P = 0.002, odds ratio [OR] = 2.524) and allele A (P = 0.0003, OR = 1.613) of the WNT2 rs4730775 polymorphism were associated with HCC susceptibility compared with healthy controls. Genotype GA (P = 0.001, OR = 0.567) and allele A (P = 0.002, OR = 0.652) of rs3864004, and genotype AG (P = 0.0004, OR = 0.495) and allele G (P = 0.001, OR = 0.596) of rs11564475 in the CTNNB1 gene were correlated with HCC compared with N-HCC patients. These findings were consistent in dominant and recessive models. Multidimensionality reduction analysis revealed that interactions among rs3864004, rs11564475, and rs4730775 were significantly associated with HCC compared with N-HCC patients. The polymorphism rs4135385 of CTNNB1 genotype GA was associated with a higher risk for Stage III + IV HCC (modified Union for International Cancer Control) (P = 0.001, OR = 2.238).Genetic polymorphisms in the WNT2 and CTNNB1 genes were closely associated with HCC risk and progression in a Chinese Han population.

ADP-glucose pyrophosphorylase gene plays a key role in the quality of corm and yield of cormels in gladiolus.

Starch is the main storage compound in underground organs like corms. ADP-glucose pyrophosphorylase (AGPase) plays a key role in regulating starch biosynthesis in storage organs and is likely one of the most important determinant of sink strength. Here, we identify an AGPase gene (GhAGPS1) from gladiolus. The highest transcriptional levels of GhAGPS1 were observed in cormels and corms. Transformation of GhAGPS1 into Arabidopsis rescued the phenotype of aps1 mutant. Silencing GhAGPS1 in gladiolus corms by virus-induced gene silencing (VIGS) decreased the transcriptional levels of two genes and starch content. Transmission electron microscopy analyses of leaf and corm sections confirmed that starch biosynthesis was inhibited. Corm weight and cormel number reduced significantly in the silenced plants. Taken together, these results indicate that inhibiting the expression of AGPase gene could impair starch synthesis, which results in the lowered corm quality and cormel yield in gladiolus.

[Medical negligence in surgery: 112 cases retrospective analysis].

OBJECTIVE: To explore the general characteristics of medical negligence in surgery in order to provide the reference for forensic practices. METHODS: One hundred and twelve cases of medical negligence in surgical department were retrospectively analyzed in Fada Institute of Forensic Medicine and Science from 2008 to 2010. RESULTS: The common types of medical negligence cases in the surgery were improper operation procedure (28.57%), failure of consent (26.79%), and inadequate monitoring (22.32%). The results of complications included disability or functional impairment (61.61%), death (31.25%) and transient impairment with no obvious adverse reactions (7.14%). The most common roles played by the medical negligence cases were minor role (26.79%), equal role (19.64%), and slight role (14.29%). CONCLUSION: Significant attention should be paid to the operation procedure, consent, and monitoring. It should be cautious to not make assessment on involvement degree of medical negligence.

A novel metallothionein gene from a mangrove plant Kandelia candel.

A new metallothionein (MT) gene was cloned from Kandelia candel, a mangrove plant with constitutional tolerance to heavy metals, by rapid amplification of cDNA ends and named KMT, which is composed of two exons and one intron. The full length of KMT cDNA was 728 bp including 121 bp 5' noncoding domain, 240 bp open reading frame and 384 bp 3' termination. The coding region of KMT represented a putative 79 amino acid protein with a molecular weight of 7.75 kDa. At each of the amino- and carboxy-terminal of the putative protein, cysteine residues were arranged in Cys-Cys, Cys-X-Cys and Cys-X-X-Cys, indicating that the putative protein was a novel type 2 MT. Sequence and homology analysis showed the KMT protein sequence shared more than 60 % homology with other plant type 2 MT-like protein genes. At amino acid level, the KMT was shown homology with the MT of Quercus suber (83 %), of Ricinus communis (81 %) and of Arabidopsis thaliana (64 %). Function studies using protease-deficient Escherichia coli strain BL21 Star ™(DE3) confirmed the functional nature of this KMT gene in sequestering both essential (Zn) and non-essential metals (Cd and Hg) and the E. coli BL21 with KMT can live in 1,000 µmol/L Zn, 120 µmol/L Hg, and 2,000 µmol/L Cd. The information could provide more details of the causative molecular and biochemical mechanisms (including heavy metal sequestration) of the KMT in K. candel or a scientific basis for marine heavy-metal environment remediation with K. candel. This study also provides a great significance of protecting mangrove species and mangrove ecosystem.

[Analysis of autopsy and clinical findings on medical malpractice cases after heart operation].

OBJECTIVE: The purpose of this study was to use autopsy to explore cause of death and to identify medical errors after cardiac surgery. METHODS: Clinical and autopsy findings in 6 cases were analyzed with respect to the clinical diagnosis, operation types, death time and features, and autopsy findings, medico-legal disputes and related medical errors. RESULTS: There were total 6 patients. The procedures involve cardiac valve replacement (4), coronary artery bypass (1), and congenital aortic transposition repair (1). Three patients had sudden death one week after surgery and 3 from congestive heart failure. The findings include myocardial infarction (2), massive myocardial injury (1), endocarditis (2), and multi-organ failure (1). The families in all six cases suspected malpractice. The major concerns were operation indication and timing, selection of operation equipment, operative mishandling, inadequate post-operative care and timely therapeutic invention, inadequate informed consent regarding the severity of the disease itself, the risks of heart surgery, and its prognosis after the procedures. CONCLUSION: Autopsy can be used to determine the cause of death, to assess the quality of the operation and post operation management, and to help to resolve malpractice disputes

Differential regulation of the human kappa opioid receptor by agonists: etorphine and levorphanol reduced dynorphin A- and U50,488H-induced internalization and phosphorylation.

We previously observed that (trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]benzeneacetamide (U50,488H) promoted internalization and phosphorylation of the FLAG-tagged human kappa opioid receptor (FLAG-hkor) stably expressed in Chinese hamster ovary (CHO) cells. In this study, we compared regulation of the FLAG-hkor expressed in CHO cells by U50,488H, dynorphin A, etorphine, and levorphanol, which were potent full agonists as determined by stimulation of guanosine 5'-O-(3-[(35)S]thio)triphosphate binding. Using fluorescence flow cytometry, we found that dynorphin A(1-17), like U50,488H, promoted internalization of the FLAG-hkor in a time- and dose-dependent manner. The antagonists naloxone and norbinaltorphimine, having no effect on FLAG-hkor internalization, effectively blocked dynorphin A(1-17)- and U50,488H-induced internalization. Interestingly, the full agonists etorphine and levorphanol did not cause internalization of the FLAG-hkor but significantly reduced dynorphin A(1-17)- and U50,488H-induced internalization in a dose-dependent manner. Immunofluorescence staining of FLAG-hkor yielded similar results. Dynorphin A(1-17) and U50,488H enhanced phosphorylation of FLAG-hkor to a greater extent than etorphine, but levorphanol did not increase FLAG-hkor phosphorylation. Etorphine or levorphanol decreased dynorphin- or U50,488H-induced phosphorylation. It is likely that conformations of the hkor required for phosphorylation and initiation of internalization are different from those for activation of G proteins. We also examined whether the four agonists had differential effects on superactivation of adenylate cyclase. Pretreatment with U50,488H, dynorphin A(1-17), or etorphine enhanced forskolin-stimulated adenylate cyclase activity to approximately 200 to 250% of the control, whereas levorphanol pretreatment did not result in significant adenylate cyclase superactivation. Thus, the degree of superactivation caused by an agonist is unrelated to its ability to promote internalization of the hkor.

Molecular basis of differences in (-)(trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidiny)-cyclohexyl]benzeneacetamide-induced desensitization and phosphorylation between human and rat kappa-opioid receptors expressed in Chinese hamster ovary cells.

The agonist (-)(trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidiny)-cyclohexyl]benzeneacetamide [(-)U50,488H] caused desensitization of the human kappa-opioid receptor (hkor) and Flag-tagged hkor (Flag-hkor) but not the rat kappa-opioid receptor (rkor) and Flag-tagged rkor (Flag-rkor) stably expressed in CHO cells as assessed by guanosine 5'-O-(3-[35S]thiotriphosphate) binding. In addition, (-)U50,488H stimulation enhanced phosphorylation of the Flag-hkor, but not Flag-rkor. (-)U50,488H-induced phosphorylation of the Flag-hkor was reduced by expression of the dominant negative mutant GRK2-K220R, demonstrating the involvement of G protein-coupled receptor kinases (GRKs). However, expression of GRK2 and arrestin-2 or GRK3 and arrestin-3 did not result in desensitization or phosphorylation of the Flag-rkor after (-)U50,488H pretreatment. To understand the molecular basis of the species differences, we constructed two Flag-tagged chimeric receptors, Flag-h/rkor and Flag-r/hkor, in which the C-terminal domains of Flag-hkor and Flag-rkor were switched. When stably expressed in CHO cells, Flag-r/hkor, but not Flag-h/rkor, was desensitized and phosphorylated after exposure to (-)U50,488H, indicating that the C-terminal domain plays a critical role in the differences. We then generated a Flag-hkor mutant, in which S358 was mutated to N (Flag-hkorS358N) and a Flag-rkor mutant, in which N358 was substituted with S (Flag-rkorN358S). Although Flag-hkorS358N was not phosphorylated or desensitized by (-)U50,488H stimulation, Flag-rkorN358S underwent (-)U50,488H-induced desensitization with slightly increased phosphorylation. These results indicate that there are differences in (-)U50,488H-induced desensitization and phosphorylation between the hkor and the rkor. In addition, the C-terminal domain plays a crucial role in these differences and the 358 locus contributes to the differences. Our findings suggest caution in extrapolating studies on kappa-opioid receptor regulation from rats to humans.