Fabrication of levofloxacin-loaded porcine acellular dermal matrix hydrogel and functional assessment in urinary tract infection.

Bacterial cystitis, a commonly occurring urinary tract infection (UTI), is renowned for its extensive prevalence and tendency to recur. Despite the extensive utilization of levofloxacin as a conventional therapeutic approach for bacterial cystitis, its effectiveness is impeded by adverse toxic effects, drug resistance concerns, and its influence on the gut microbiota. This study introduces Lev@PADM, a hydrogel with antibacterial properties that demonstrates efficacy in the treatment of bacterial cystitis. Lev@PADM is produced by combining levofloxacin with decellularized porcine acellular dermal matrix hydrogel and exhibits remarkable biocompatibility. Lev@PADM demonstrates excellent stability as a hydrogel at body temperature, enabling direct administration to the site of infection through intravesical injection. This localized delivery route circumvents the systemic circulation of levofloxacin, resulting in a swift and substantial elevation of the antimicrobial agent's concentration specifically at the site of infection. The in vivo experimental findings provide evidence that Lev@PADM effectively prolongs the duration of levofloxacin's action, impedes the retention and invasion of E.coli in the urinary tract, diminishes the infiltration of innate immune cells into infected tissues, and simultaneously preserves the composition of the intestinal microbiota. These results indicate that, in comparison to the exclusive administration of levofloxacin, Lev@PADM offers notable benefits in terms of preserving the integrity of the bladder epithelial barrier and suppressing the recurrence of urinary tract infections.

Synovial sarcoma X breakpoint 1 protein uses a cryptic groove to selectively recognize H2AK119Ub nucleosomes.

The cancer-specific fusion oncoprotein SS18-SSX1 disturbs chromatin accessibility by hijacking the BAF complex from the promoters and enhancers to the Polycomb-repressed chromatin regions. This process relies on the selective recognition of H2AK119Ub nucleosomes by synovial sarcoma X breakpoint 1 (SSX1). However, the mechanism underlying the selective recognition of H2AK119Ub nucleosomes by SSX1 in the absence of ubiquitin (Ub)-binding capacity remains unknown. Here we report the cryo-EM structure of SSX1 bound to H2AK119Ub nucleosomes at 3.1-Å resolution. Combined in vitro biochemical and cellular assays revealed that the Ub recognition by SSX1 is unique and depends on a cryptic basic groove formed by H3 and the Ub motif on the H2AK119 site. Moreover, this unorthodox binding mode of SSX1 induces DNA unwrapping at the entry/exit sites. Together, our results describe a unique mode of site-specific ubiquitinated nucleosome recognition that underlies the specific hijacking of the BAF complex to Polycomb regions by SS18-SSX1 in synovial sarcoma.

Mechanistic insights into nucleosomal H2B monoubiquitylation mediated by yeast Bre1-Rad6 and its human homolog RNF20/RNF40-hRAD6A.

Histone H2B monoubiquitylation plays essential roles in chromatin-based transcriptional processes. A RING-type E3 ligase (yeast Bre1 or human RNF20/RNF40) and an E2 ubiquitin-conjugating enzyme (yeast Rad6 or human hRAD6A), together, precisely deposit ubiquitin on H2B K123 in yeast or K120 in humans. Here, we developed a chemical trapping strategy and successfully captured the transient structures of Bre1- or RNF20/RNF40-mediated ubiquitin transfer from Rad6 or hRAD6A to nucleosomal H2B. Our structures show that Bre1 and RNF40 directly bind nucleosomal DNA, exhibiting a conserved E3/E2/nucleosome interaction pattern from yeast to humans for H2B monoubiquitylation. We also find an uncanonical non-hydrophobic contact in the Bre1 RING-Rad6 interface, which positions Rad6 directly above the target H2B lysine residue. Our study provides mechanistic insights into the site-specific monoubiquitylation of H2B, reveals a critical role of nucleosomal DNA in mediating E3 ligase recognition, and provides a framework for understanding the cancer-driving mutations of RNF20/RNF40.

An-Luo-Hua-Xian Pill Improves the Regression of Liver Fibrosis in Chronic Hepatitis B Patients Treated with Entecavir.

Background and Aims: Chronic hepatitis B (CHB) can cause liver fibrosis and lead to cirrhosis and cancer. As the effectiveness of antiviral therapy to reverse liver fibrosis is limited, We aimed to evaluate the effect of An-Luo-Hua-Xian pill (ALHX) on fibrosis regression in CHB patients treated with entecavir (ETV). Methods: Treatment-naïve patients with CHB were randomly treated with ETV alone or combined with ALHX (ETV+ALHX) between October 1, 2013 and December 31, 2020. Demographic, laboratory, and liver histology data before and after 78 weeks of treatment were collected. The Ishak fibrosis score (F) was used and fibrosis regression required a decrease in F of ≥1 after treatment. Results: A total of 780 patients were enrolled, and 394 with a second liver biopsy after treatment were included in the per-protocol population, 132 in ETV group and 262 in ETV+ALHX group. After 78 weeks of treatment, the fibrosis regression rate in the ETV+ALHX group was significantly higher than that of the ETV group at baseline F≥3 patients: 124/211 (58.8%) vs. 45/98 (45.9%), p=0.035. The percentage of patients with a decreased liver stiffness measurement (LSM) was higher in the ETV+ALHX group: 156/211 (73.9%) vs. 62/98 (63.%), p=0.056. Logistic regression analysis showed that ETV combined with ALHX was associated with fibrosis regression [odds ratio (OR)=1.94, p=0.018], and a family history of hepatocellular carcinoma was on the contrary. (OR=0.41, p=0.031). Conclusions: ETV combined with ALHX increased liver fibrosis regression in CHB patients.

The median effective volume of ultrasound-guided thoracic paravertebral nerve block with 0.3% ropivacaine in radical thoracoscopic surgery for lung cancer.

BACKGROUND: Ultrasound-guided needle placement has revolutionized the thoracic paravertebral block technique and can be applied in thoracoscopic surgery. OBJECTIVE: This study investigated the median effective volume (EV50) of an ultrasound-guided single shot of 0.3% ropivacaine used as a thoracic paravertebral nerve block for the radical thoracoscopic resection of lung cancer. METHODS: A total of 27 patients who received a single shot of ultrasound-guided thoracic paravertebral nerve block and underwent radical thoracoscopic resection of lung cancer were enrolled in this study between February 10 and August 13, 2018. All patients were rated as ASA grades I or II. Using ultrasound as a guide, the block needle was gradually pushed through the lateral costotransverse ligaments to the thoracic paravertebral space by the in-plane technique. After confirming the absence of blood or cerebrospinal fluid, 1-2 ml of 0.3% ropivacaine hydrochloride was injected to confirm that the position of the needle was appropriate, and a pre-determined volume of 0.3% ropivacaine hydrochloride was then administered to the patients. Sensory testing by pinprick was performed every 5 minutes for 30 minutes following the thoracic paravertebral block injection to identify the time segments during which the loss of sensation to the pinprick and its blocking effect occurred. RESULTS: All patients completed the study and 14 (51.8%) had a successful block. CONCLUSION: The EV50 of 0.3% ropivacaine was 18.46 ml (95% CI 17.09-19.95 ml) and the EV95 was 20.89 ml.

Malignant melanoma mimic fungal infection a case report.

BACKGROUND: Most of malignant melanomas originate from skin and often metastasize to the lungs, rarely metastasizes to the liver and bone. However, imageology characters of lung metastasis tumor are commonly similar to those of fungal infections. CASE PRESENTATION: A patient was admitted with unhealed plantar puncture wound for 3 years, and cough and expectoration for 2 years. The chest computed tomography (CT) revealed multiple nodules with cavities, and the patient was diagnosed of pulmonary fungal infection in another hospital and received antifungal therapy for more than 8 months, but the clinical symptoms and chest imaging findings continue to progress. After admission, the pathological results of both lung biopsy and biopsy of the plantar wound 3 years ago indicated malignant melanoma. CONCLUSIONS: The diagnosis of lung lesions cannot rely solely on imaging diagnosis, lung biopsy should be performed if necessary.

Chemical methods for studying the crosstalk between histone H2B ubiquitylation and H3 methylation.

The reversible and dynamic post-translational modifications (PTMs) of histones in eukaryotic chromatin are intimately connected to cell development and gene function, and abnormal regulation of PTMs can result in cancer and neurodegenerative diseases. Specific combinations of these modifications are mediated by a series of chromatin proteins that write, erase, and read the "histone codes," but mechanistic studies of the precise biochemical and structural relationships between different sets of modifications and their effects on chromatin function constitute a unique challenge to canonical biochemical approaches. In the past decade, the development and application of chemical methods for investigating histone PTM crosstalks has received considerable attention in the field of chemical biology. In this review, taking the functional crosstalk between H2B ubiquitylation at Lys120 (H2BK120ub) and H3 methylation at Lys79 (H3K79me) as a typical example, we survey recent developments of different chemical methods, in particular, protein synthetic chemistry and protein-based chemical probes, for studying the mechanism of the functional crosstalks of histone PTMs.

Quantitative proteomic analysis of glycosylated proteins enriched from urine samples with magnetic ConA nanoparticles identifies potential biomarkers for small cell lung cancer.

Lung cancer has high morbidity and mortality and small cell lung cancer (SCLC) is a highly invasive malignant tumor with a very unfavorable survival rate. Early diagnosis and treatment can result in better prognosis for the SCLC patients but current diagnostic methods are either invasive or incapable for large-scale screen. Therefore, discovering biomarkers for early diagnosis of SCLC is of importance. In this work, we covalently coupled Concanavalin A (ConA) to functionalized magnetic nanoparticles to obtain magnetic ConA-nanoparticles (ConA-NPs) for the enrichment of glycosylated proteins. We then purified glycosylated proteins in 36 urine samples from 9 healthy controls, 9 SCLC patients, 9 lung adenocarcinoma (LUAD) patients, and 9 lung squamous cell carcinoma (LUSC) patients. The purified glycosylated proteins were digested and analyzed by LC-MS/MS for identification and quantification. Among the 398 identified proteins, 20, 15, and 1 glycosylated protein(s), respectively, were upregulated in the urine of SCLC, LUAD, and LUSC patients. Immunoblotting experiments further demonstrated that cathepsin C and transferrin were significantly upregulated in the ConA-NP purified urine of SCLC patients. This work suggests that glycosylated cathepsin C and transferrin might be able to serve as potential biomarkers for the noninvasive diagnosis of SCLC patients.

A Comparison of Puncture and Continuous Pump Analgesia With Two Different Approaches to Thoracic Paravertebral Block for Thoracic Surgery.

BACKGROUND: This study aims to compare the success rate of thoracic paravertebral block (TPVB) and the effect of postoperative analgesia between two approaches. METHODS: A total of 34 patients with American Society of Anesthesiology (ASA) physical status score II-III, undergoing an optional thoracoscopic surgery, were randomly assigned to a parasagittal approach group (group P, n = 17) and a transverse intercostal approach group (group T, n = 17). The catheterization time, success rate of the puncture and catheterization, block plane and effect at the surgical site were compared between two groups. The mean arterial pressure and heart rate were recorded, as well as the cold tactile block plane and numeric rating scale (NRS) at 0.5, 2, 4, 8, 12, 24, and 48 h after surgery. The study was registered at http://www.chictr.org.cn/showproj.aspx?proj=9624 (Registration number: ChiCTR2100054642). RESULTS: The catheterization time in group P was significantly longer than that in group T (P < 0.05). The success rate of catheterization in group P was lower than that in group T, but no statistical significance (P = 0.085). There was no significant difference in the success rate of Puncture and blocking effect of the surgical site at 30 min post-injection between two groups (P > 0.05). There was no significant difference in the cold tactile block plane and NRS scores during coughing between two groups at 0.5, 2, 4, 8, 12, 24, and 48 h postoperatively (P > 0.05). CONCLUSION: This study suggests that there is no significant difference in postoperative block level or pain score during coughing for thoracoscopic surgery between ultrasound-guided parasagittal and transverse intercostal approach, but the parasagittal approach takes longer and has a higher failure rate.

A Comparison of Analgesia After a Thoracoscopic Lung Cancer Operation with a Sustained Epidural Block and a Sustained Paravertebral Block: A Randomized Controlled Study.

INTRODUCTION: This study aimed to compare the challenge of puncture and catheterization and the effect of postoperative analgesia of ultrasound-guided continuous thoracic paravertebral block and the continuous epidural analgesia in patients receiving thoracoscopic surgery for lung cancer. METHODS: One-hundred and fifty patients received elective unilateral thoracoscopic surgery for lung cancer and were randomized into three groups; test group 1 (T group), test group 2 (P group), and the control group (E group). Both of the test groups received ultrasound-guided continuous thoracic paravertebral block (TPVB) while the control group received continuous epidural analgesia. After the operation, all the patients in the test groups received the same postoperative analgesia; loading dose 0.5 mg kg-1, background dose 0.25 mg kg-1 h-1, patient controlled analgesia (PCA) 0.25 mg kg-1, and a locking time of 60 min, while the patients in the control group received a loading dose of 5 ml, a background dose of 5 ml h-1, and a locking time of 20 min. The outcomes of this study were the success rate of the puncture and catheter placement, the blocked segments, numerical rating scale (NRS) scores at rest and during coughing, and the segments with reduced or lost cold and pinpricking sensation. RESULTS: The success rates of the puncture and catheterization in group T were the highest. Compared with group P, the failure rate of the puncture in group E was lower (p < 0.05), but the success rate of catheterization was higher (p < 0.05). The puncture time in group T was the shortest; there was no difference between group E and group T. The time of catheterization in group P was the longest, this was followed by group T, and was the fastest in group E. The stable time of the block level in group E was shorter than that in groups P and T, but was similar between groups P and T. The block level of all three groups in the 4 h postoperative period was similar (p > 0.05), while the 4 h postoperative levels of groups P and T were reduced significantly (p < 0.05). CONCLUSION: The continuous analgesia technique of paravertebral space catheterization cannot replace the continuous epidural analgesia in thoracoscopic lung cancer surgery as the latter technique is still considered to be the gold standard. TRIAL REGISTRATION: China Clinical Trial Registration Center identifier ChiCTR1900020973.

Significance of 125I radioactive seed implantation on growth differentiation factor and programmed death receptor-1 during treatment of oral cancer.

BACKGROUND: Oral cancer (OC) is the most common malignant tumor in the oral cavity, and is mainly seen in middle-aged and elderly men. At present, OC is mainly treated clinically by surgery or combined with radiotherapy and chemotherapy; but recently, more and more studies have shown that the stress trauma caused by surgery and the side effects of radiotherapy and chemotherapy seriously affect the prognosis of patients. AIM: To determine the significance of 125I radioactive seed implantation on growth differentiation factor 11 (GDF11) and programmed death receptor-1 (PD-1) during treatment of OC. METHODS: A total of 184 OC patients admitted to The Second Affiliated Hospital of Jiamusi University from May 2015 to May 2017 were selected as the research subjects for prospective analysis. Of these patients, 89 who received 125I radioactive seed implantation therapy were regarded as the research group (RG) and 95 patients who received surgical treatment were regarded as the control group (CG). The clinical efficacy, incidence of adverse reactions and changes in GDF11 and PD-1 before treatment (T0), 2 wk after treatment (T1), 4 wk after treatment (T2) and 6 wk after treatment (T3) were compared between the two groups. RESULTS: The efficacy and recurrence rate in the RG were better than those in the CG (P < 0.05), while the incidence of adverse reactions and survival rate were not different. There was no difference in GDF11 and PD-1 between the two groups at T0 and T1, but these factors were lower in the RG than in the CG at T2 and T3 (P < 0.05). Using receiver operating characteristic (ROC) curve analysis, GDF11 and PD-1 had good predictive value for efficacy and recurrence (P < 0.001). CONCLUSION: 125I radioactive seed implantation has clinical efficacy and can reduce the recurrence rate in patients with OC. This therapy has marked potential in clinical application. The detection of GDF11 and PD-1 in patients during treatment showed good predictive value for treatment efficacy and recurrence in OC patients, and may be potential targets for future OC treatment.

Sensitive and selective SERS probe for detecting the activity of γ-glutamyl transpeptidase in serum.

γ-Glutamyl transpeptidase (GGT) has attracted considerable attention for its regulatory effect on glutathione metabolism in living organisms; further, its close relationship with physiological dysfunctions such as hepatitis and liver cancers has enhanced its applicability. Therefore, the accurate detection of GGT levels is particularly important for the early diagnosis of diseases. Thus, we herein report the development of a surface-enhanced Raman spectroscopic (SERS) probe, namely bis-s,s'-((s)-4,4'-thiolphenylamide-Glu) (b-(s)-TPA-Glu), that comprises of a γ-glutamyl moiety for detection of the GGT activity. In this system, detection was achieved by observing differences in the SERS spectral profiles of the b-(s)-TPA-Glu probe and its corresponding hydrolysis product that resulted from the catalytic action of GGT. This SERS probe system exhibited a high selectivity toward GGT due to a combination of its specific catalytic action and the distinctive spectroscopic fingerprint of the SERS technique. The developed SERS approach was also found to be approximately linear in the range of 0.2-200 U/L, and a limit of detection of 0.09 U/L was determined. Furthermore, the proposed SERS method was suitable for detection of the GGT activity of clinical serum samples and also for evaluation of the inhibitors of GGT. Consequently, this approach is considered to be a promising diagnostic and drug screening tool for GGT-associated diseases.

Thiirane linkers directed histone H2A diubiquitination suggests plasticity in 53BP1 recognition.

Polyubiquitination with diverse linkages on histones provides another layer of accuracy and complexity for epigenetic regulation, which is rarely studied. Herein, K27 or K48-diubiquitin modified H2A analogues were chemically synthesized using thiirane linkers. These permitted in vitro binding studies suggested the plasticity of ubiquitin chains in 53BP1 recognition.

Candida albicans-induced acute lung injury through activating several inflammatory signaling pathways in mice.

Candida albicans infection-induced acute lung injury is one of the most prevalent diseases in immunosuppressive individual. Nevertheless, the mechanism by which Candida albicans induced acute lung injury remains unclear. The present study investigated the mechanism by which Candida albicans induced acute lung injury in mice. Mice were randomly divided into four groups and intratracheally injected with 60 µl Candida albicans (106 CFU) or normal saline. Half of the mice were sacrificed at 6 h after Candida albicans. The rest of the mice for survival test were observed until 7 d after Candida albicans. As expected, immunosuppression aggravated Candida albicans-induced acute lung injury and death in mice. Additionally, Candida albicans infection elevated mRNA levels of pro-inflammatory and chemokines in lungs and the levels of IL-6, IL-1ß and IL-17 in serum. Further study showed that Candida albicans promoted nuclear translocation of NF-κB p50 and p65 subunits in pulmonary epithelial cells and interstitial cells. Candida albicans induced pulmonary p38, ERK1/2 and Akt phosphorylation in normal and immunosuppressive mice. Moreover, Candida albicans infection activated pulmonary STAT3 signaling in normal and immunosuppressive mice. Overall, these results suggest that Candida albicans induced acute lung injury and death may be through activating several inflammatory signaling pathways.

Obeticholic acid alleviate lipopolysaccharide-induced acute lung injury via its anti-inflammatory effects in mice.

Acute lung injury (ALI) is a common disease that may result in acute respiratory failure and death. However, there are still no effective treatments for ALI. Several studies have shown that farnesoid X receptor (FXR) has an anti-inflammatory effect. We investigated the effects of obeticholic acid (OCA), an agonist of FXR, on Lipopolysaccharide (LPS)-induced ALI in mice. Sixty male mice were randomly divided into six groups, and orally administered with or without OCA once daily for 3 consecutive days before LPS (1.0 mg/kg). Animals were sacrificed at 0 h, 2 h or 6 h after LPS. As expected, OCA enhanced pulmonary FXR activity. OCA prevented LPS-induced ALI. Additional experiment showed that OCA alleviated LPS-induced up-regulation of pulmonary pro-inflammatory and chemokine genes. Moreover, OCA also repressed LPS-induced the release of TNF-α and KC in serum and bronchoalveolar lavage fluid. In contrast, OCA further up-regulated LPS-induced the expression of Il-10, an anti-inflammatory cytokine. Further study showed that OCA inhibited LPS-evoked NF-κB signaling in the lungs. OCA attenuated LPS-induced ERK1/2, JNK, p38 and Akt phosphorylation in the lungs. Overall, these results suggest that OCA prevent LPS-induced ALI may be through enhancing pulmonary FXR activity and then blockading several inflammatory signaling pathways.

Clinical application and effect of dexmedetomidine in combination with continuous positive airway pressure on one-lung ventilation in lung surgery of elder patients.

In this study, 126 elder patients who underwent thoracic surgery under general anesthesia were divided into three groups randomly, i.e. dexmedetomidine group, positive ventilation group and combination group. All patients received varying strategies in addition to the one-lung ventilation, and changes in oxidative stress and indicators of inflammation at different time points were observed. In comparison to the dexmedetomidine group and the positive ventilation group, patients in the combination group at T2-3 had lower levels of malonaldehyde, cortisol, C-reaction protein, interleukin-6 (IL-6) and tumor necrosis factor α in serum (p<0.05); one day after surgery, the incidence of complications in lungs of patients in the combination group was significantly lower than those in the dexmedetomidine group and the positive ventilation group (p<0.05). Dexmedetomidine in combination with continuous positive airway pressure can alleviate the oxidative stress and inflammation of lung tissues in one-lung ventilation during the thoracic surgery of elder patients, thus, reducing the incidence of postoperative complications.

[General situation and comprehensive utilization of medicinal fungi resources in Heilongjiang province].

In order to develop and utilize the macrofungi in Heilongjiang province, numerous literatures have been investigated to make a comprehensive analysis of the number of known species of fungi in Heilongjiang province. There exists a total of 546 species of macrofungus in Heilongjiang province belonging to 53 families and 13 orders of 6 classes and 2 subdivisions. And its application value is classified, summarized and reviewed. Three hundred and twenty kinds of edible fungi, 214 species of fungi with medicinal value, medicinal value in the anti-cancer effects of 167 species of fungi, 141 wood rot fungi, 141 species of ectomycorrhizal fungi, 88 poisonous species, 67 macrofungus which are not clarified whether could be edible or toxic. It shows a broad prospects for development and utilization of macrofungus resources in Heilongjiang province.

Predict esophageal varices via routine trans-abdominal ultrasound: A design of classification analysis model.

BACKGROUND AND AIMS: Upper gastrointestinal endoscopy remains the gold standard for diagnosis of esophageal varices. Trans-abdominal ultrasound, as a noninvasive routine examination for the follow-up of cirrhosis patient, is safe, cheap, easy to perform, and plays an important role. In this study, we attempt to design a practical classification analysis model to predict esophageal varices via ultrasound. METHODS: Compared with endoscopy, the ultrasound qualitative signs (lower esophageal Doppler signals, left gastric vein hepatofugal flow, and paraumbilical vein recanalization) and quantitative parameters (spleen diameter, spleen vein diameter, portal vein diameter, and portal vein velocity) have been evaluated in 286 cirrhosis patients. RESULTS: The classification analysis model is designed as that: the patients are defined with esophageal varices high risk, who with any ultrasound qualitative signs or who with spleen diameter greater than 162 mm without qualitative parameters. The sensitivity for detecting esophageal varices is 97.5% and the specificity is 82.6%, while the positive predictive value is 96.7%, negative predictive value is 83.4%, and the omission diagnostic rate is 2.5%. CONCLUSIONS: This classification analysis model design includes ultrasound qualitative signs and spleen diameter, which can be detected easily via routine ultrasound without other auxiliary. The classification analysis model is useful in detecting esophageal varices, which may be a supplement for predicting of esophageal varices, and reducing the frequency of endoscopy in the follow-up of cirrhosis patients.