OBJECTIVETo investigate histone acetylation modification of topoisomerase enzyme Ⅱα (TOPOⅡα) promoter regulation factors in patients with chronic benzene poisoning, to explore the possible regulatory mechanism of TOPOⅡα involved in toxicity of chronic benzene poisoning;

METHODSThe bone marrow samples were from 25 chronic benzene poisoning cases and 25 controls. The Chromatin Immunoprecipitation (ChIP) assay was carried out to study the possible mechanism of TOPOⅡα promoter regulation factors expression changes. TOPOⅡα promoter regulation factors mRNA were detected by RT-PCR technique.

RESULTS(1) Compared with the control, the histone H4 acetylation, histone H3 acetylation level of TOPOⅡα promoter regulation factors SP1, ATF-2, SP3, NF-YA, P53, C-MYB, ICBP90, NF-M in chronic benzene poisoning patients decreased, with the significant difference (P<0.05) , except for C-JUN (P>0.05) ; (2) The mRNA expression of TOPOⅡαpromoter regulation factors SP1, NF-YA, C-MYB, C-JUN and NF-M were significantly lower than in the control with the significant difference (P<0.05) , while the expression of SP3、P53 mRNA increased (P<0.05) , ATF-2、ICBP90 mRNA wasn't changed (P>0.05) .

CONCLUSION(1) Chronic benzene poisoning TOPO Ⅱα promoter regulation factors histone modification changes accompanied with mRNA level changed. (2) Histone acetylation modification of topoisomerase enzyme Ⅱα promoter regulation factors takes important role in the benezen's Hematopoietic toxicity.

Acetylation ; Antigens, Neoplasm ; metabolism ; Benzene ; poisoning ; Case-Control Studies ; Chromatin Immunoprecipitation ; Chronic Disease ; DNA Topoisomerases, Type II ; metabolism ; DNA-Binding Proteins ; metabolism ; Histones ; metabolism ; Humans ; Poisoning ; metabolism ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism

Objective: To evaluate the value of Prostate Imaging Reporting and Data System Version 2 (PI-RADS ) based biparametric magnetic resonance imaging (bpMRI) for predicting prostate biopsy results in patients with elevated prostate specific antigen (PSA). Methods: The bpMRI from 539 patients who took transperineal template saturate biopsy from January 2015 to October 2017 were assessed retrospectively. The average age was 69.5 years old (44-88 years), with tPSA level of 7.23 ng/ml (4-10 ng/ml), f/t PSA of 0.183( 0.016-0.504), PSAD of 0.126 ng/ml² ( 0.025-0.534 ng/ml²) , PV of 72.42 ml ( 18.71-199.51 ml). The age, PSA level, free/total PSA ratio, PSA density, prostate volume, and PI-RADS score of enrolled patients were analyzed for univariate analysis and their difference was compared by chi-square test, t-test. The multivariate logistic regression analysis was also performed through SPSS to select the independent risk factors for prostate cancer (PCa) and clinically significant cancer (csPCa). The receiver operating characteristic curves were also constructed to analyze the sensitivity and specificity of PI-RADS in PCa to explore the best cut-off value for the diagnosis of PCa and csPCa. Results: A total of 539 patients were included in our study with 244 cases being positive and 295 cases being negative. In patients with positive results, 59 patients were diagnosed csPCa. According to univariate analysis results, the age(P<0.001) and PI-RADS score (P<0.001) of the positive patients were higher than the negative patients, and the difference was statistically significant. The age of the csPCa patients (P=0.023), PSAD (P=0.048) and PI-RADS scores (P<0.001) were higher than those of InsPCa patients, and f/t PSA (P=0.027) was lower than that of InsPCa patients with statistically significance. Multivariate logistic regression analysis demonstrated that f /t PSA (OR=2.283, P=0.049) and PI-RADS score (OR=9.046, P<0.001) were independent risk factors for positive biopsy results, while PSAD (OR=4.54, P=0.038) and PI-RADS score (OR=8.254, P<0.001) were independent risk factor for csPCa. The Yoden index analysis of different thresholds for prostate cancer detection indicated that PI-RADS 3 was the optimal threshold for the diagnosis of PCa, and PI-RADS 4 was the optimal threshold for the diagnosis of csPCa. Based on the combination of the above factors, the positive rate of prostate cancer was relatively high in patients with PI-RADS score ≥3 and f/t PSA<0.2 , which accounted for 86.6%(181/209). In contrast, the positive rate in patients with a PI-RADS score of ≤2 and f/t PSA≥0.2 was low, which accounted for 10.7%(6/56). The positive rate of csPCa was relatively high in patients with PI-RADS score≥4 and PSAD≥0.15 ng/ml^2, which accounted for 76.0%(38/50). The positive rate of csPCa detected in patients with ≤3 and PSAD<0.15 ng/ml² was low, which accounted for 0(0/359). Conclusions PI-RADS score could be used to reduce the unnecessary prostate biopsies in patients with elevated PSA when combined with other PSA related markers. Patients with a PI-RADS score of ≤3 and a PSAD ratio <0.15 ng/ml² could avoid unnecessary biopsies.

Purpose: Little is known about the role of gut microbiota in the pathogenesis of erectile dysfunction (ED). We performed a study to compare taxonomic profiles of gut microbiota of ED and healthy males. Materials and Methods: A total of 43 ED patients and 16 healthy controls were enrolled in the study. The 5-item version of the International Index of Erectile Function (IIEF-5) with a cutoff value of 21 was used to evaluate erectile function. All participants underwent nocturnal penile tumescence and rigidity test. Samples of stool were sequenced to determine the gut microbiota. Results: We identified a distinct beta diversity of gut microbiome in ED patients by unweighted UniFrac analysis (R2=0.026, p=0.036). Linear discriminant analysis effect size (LEfse) analysis showed Actinomyces was significantly enriched, whereas Coprococcus_1, Lachnospiraceae_FCS020_group, Lactococcus, Ruminiclostridium_5, and Ruminococcaceae_UCG_002 were depleted in ED patients. Actinomyces showed a significant negative correlation with the duration of qualified erection, average maximum rigidity of tip, average maximum rigidity of base, tip tumescence activated unit (TAU), and base TAU. Coprococcus_1, Lachnospiraceae_FCS020_group, Ruminiclostridium_5, and Ruminococcaceae_UCG_002 were significantly correlated with the IIEF-5 score. Ruminiclostridium_5 and Ruminococcaceae_UCG_002 were positively related with average maximum rigidity of tip, average maximum rigidity of base, ΔTumescence of tip, and Tip TAU. Further, a random forest classifier based on the relative abundance of taxa showed good diagnostic efficacy with an area under curve of 0.72. Conclusions This pilot study identified evident alterations in the gut microbiome composition of ED patients and found Actinomyces was negatively correlated with erectile function, which may be a key pathogenic bacteria.

Objective:To investigate the value of hypocalcaemia for predicting trauma-induced coagulopathy (TIC) in elderly trauma patients.Methods:The clinical data of elderly trauma patients in emergency ICU of the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2015 to September 2020 were retrospectively analyzed, including age, sex, site of injury, injury severity score (ISS), Glasgow coma scale (GCS), admission arterial blood gas analysis (Ca 2+, K +), venous blood biochemical electrolyte (Ca 2+, K +, Na +); international normalized ratio (INR), activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), use of blood products, length of stay, length of stay in ICU, total cost, and clinical prognosis. Receiver operating characteristic (ROC) curves and multivariate logistic regression analysis were performed to estimate the contribution of hypocalcaemia to triggering TIC in elderly trauma patients. Results:Totally 371 elderly trauma patients were included with a mean age of (72.5±6.8) years, and 248 (66.8%) were male. ISS score of the TIC group was higher than that of the non-TIC group [25(20, 34) vs. 21(16, 29)]. Compared with the non-TIC group, the incidence of chest injury, abdominal injury and limb injury were significantly higher , while the incidence of head and neck injury were significantly lower in the TIC group (all P<0.05). The biochemical blood calcium in the TIC group was significantly lower than that in the non-TIC group [(1.97±0.19) mmol/L vs. (2.15±0.16) mmol/L, P<0.001], but there was no significant difference in blood gas calcium between the two groups. The APTT value of the TIC group [(47.6±21.8) s vs. (33.8±4.1) s], PT value [(18.0±3.9) s vs. (13.7±0.8) s] were significantly higher than that of the non-TIC group, and FIB level was significantly lower than that of the non-TIC group[(1.7±0.8) g/L vs. (2.8±0.9) g/L] (all P<0.01). The utilization rate of blood products and the total cost in the TIC group were higher than that in the non-TIC group, while the recovery rate in the TIC group was lower than that in the non-TIC group (69.8% vs. 86.4%, P<0.001). Multivariate logistic regression analysis showed that hypocalcaemia was an independent risk factor for TIC in elderly trauma patients ( OR=5.830, 95% CI: 3.295-10.314). The area under ROC curve of correlation between biochemical calcium and TIC was 0.779 (95% CI: 0.728-0.831). The optimal diagnostic cut-off value was 2.06 mmol/L. Conclusions:The decrease of biochemical serum calcium level is an independent risk factor for TIC in elderly trauma patients. Positive correction of TIC in elderly trauma patients contributes to continuous improvement of clinical prognosis.

Objective:To explore the risk factors for early trauma-induced coagulopathy (TIC) following severe trauma in the elderly patients.Methods:A case-control study was used to analyze the clinical data of 317 elderly patients with severe trauma admitted to Second Affiliated Hospital of Zhejiang University School of Medicine between February 2015 and November 2020. There were 212 males and 105 females, aged 65-96 years [(72.6±6.8)years]. The patients were divided into TIC group ( n=32) and non-TIC group ( n=285) using the international normalised ratio (INR)>1.5 as the reference standard. Sex, age, trauma sites, injury severity score (ISS), Glasgow coma scale (GCS), first body temperature on admission, shock index(SI), first laboratory results of arterial blood gas, routine blood and coagulation, blood transfusion, usage of blood product, hospitalization days and clinical outcomes were compared between the two groups. Univariate and multivariate Logistic regression analysis were used to identify the risk factors for early TIC in patients with severe trauma. Results:Differences in sex, age, injuries to the face, chest and abdomen, GCS, first body temperature and hospitalization days were not statistically significant between the two groups (all P>0.05). The two groups showed statistical differences in the ratio of injuries to head, neck and extremities, ISS, SI, pH value, base excess (BE), lactate, hemoglobin (Hb), platelet (PLT) count (first detection, lowest level), activated partial thromboplastin time (APTT), thrombin time (TT), plasma fibrinogen (FIB), blood transfusion and usage of blood product and clinical outcomes (all P<0.05). According to the univariate analysis, injuries to the head, neck and extremities, ISS, first body temperature, SI, pH value, BE, lactate, Hb, PLT, APTT, TT and FIB were correlated with the occurrence of early TIC (all P<0.05). Multiple Logistic regressions analysis showed that SI ( OR=1.54, 95% CI 1.10-2.17, P<0.05), PLT ( OR=0.67, 95% CI 0.49-0.91, P<0.05) and FIB ( OR=0.56, 95% CI 0.40-0.78, P<0.01) were significantly correlated with the occurrence of early TIC. Conclusion:For elderly patients with severe trauma, higher SI, lower PLT count and lower concentration of FIB are independent risk factors for the incidence of early TIC.

“Cold-dampness entering ying （营）” is the key to the worsening of cold-dampness epidemic， and is more common in the elderly or critically ill cases of cold-dampness epidemic with pathogen exuberance and healthy qi deficiency. This paper reported a case of critically ill COVID-19 combined with multiple organ dysfunction treated by integrative traditional Chinese and western medicine based on “cold-dampness entering ying” theory. The patient did not have high fever after being infected with SARS-Cov-2， but D-dimer continued to increase， and she developed multiple thrombosis throughout the body and multiple organ dysfunctions such as pulmonary embolism， edema， oliguria， and shock. The patient were with enlarged and dusky tongue， with yellow， thick and greasy coating， and sublingual blood stasis， and thready， rapid and rough pulse. All these were characteristic manifestations of “cold-dampness entering ying”， and was differentiated as cold-dampness stasis. For the treatment， symptomatic and supportive western medicine of improving heart function， anti-infection， relieving asthma， stopping cough and reducing phlegm was given as the basic therapy， and additionally， traditional Chinese medicine to open the constraint and the blocked， save from collapse and restore yang， boost qi and relieve collapse， invigorate blood and drain water was used， usually with Modified Poge Zilong Xuanbai Chengqi Decoction （破格子龙宣白承气汤加减）， which was in accordance with the pathogenesis and thus achieving good effect.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease affecting both upper and lower motor neurons (MNs) with large unmet medical needs. Multiple pathological mechanisms are considered to contribute to the progression of ALS, including neuronal oxidative stress and mitochondrial dysfunction. Honokiol (HNK) has been reported to exert therapeutic effects in several neurologic disease models including ischemia stroke, Alzheimer's disease and Parkinson's disease. Here we found that honokiol also exhibited protective effects in ALS disease models both in vitro and in vivo. Honokiol improved the viability of NSC-34 motor neuron-like cells that expressed the mutant G93A SOD1 proteins (SOD1-G93A cells for short). Mechanistical studies revealed that honokiol alleviated cellular oxidative stress by enhancing glutathione (GSH) synthesis and activating the nuclear factor erythroid 2-related factor 2 (NRF2)-antioxidant response element (ARE) pathway. Also, honokiol improved both mitochondrial function and morphology via fine-tuning mitochondrial dynamics in SOD1-G93A cells. Importantly, honokiol extended the lifespan of the SOD1-G93A transgenic mice and improved the motor function. The improvement of antioxidant capacity and mitochondrial function was further confirmed in the spinal cord and gastrocnemius muscle in mice. Overall, honokiol showed promising preclinical potential as a multiple target drug for ALS treatment.

Bibenzyls, a kind of important plant polyphenols, have attracted growing attention for their broad and remarkable pharmacological activities. However, due to the low abundance in nature, uncontrollable and environmentally unfriendly chemical synthesis processes, these compounds are not readily accessible. Herein, one high-yield bibenzyl backbone-producing Escherichia coli strain was constructed by using a highly active and substrate-promiscuous bibenzyl synthase identified from Dendrobium officinale in combination with starter and extender biosynthetic enzymes. Three types of efficiently post-modifying modular strains were engineered by employing methyltransferases, prenyltransferase, and glycosyltransferase with high activity and substrate tolerance together with their corresponding donor biosynthetic modules. Structurally different bibenzyl derivatives were tandemly and/or divergently synthesized by co-culture engineering in various combination modes. Especially, a prenylated bibenzyl derivative ( 12) was found to be an antioxidant that exhibited potent neuroprotective activity in the cellular and rat models of ischemia stroke. RNA-seq, quantitative RT-PCR, and Western-blot analysis demonstrated that 12 could up-regulate the expression level of an apoptosis-inducing factor, mitochondria associated 3 (Aifm3), suggesting that Aifm3 might be a new target in ischemic stroke therapy. This study provides a flexible plug-and-play strategy for the easy-to-implement synthesis of structurally diverse bibenzyls through a modular co-culture engineering pipeline for drug discovery.