Optimizing lung biopsy procedures:Comparative analysis of diagnostic efficacy and safety in experimental low-dose, conventional low-dose, and standard-dose CT-guided approaches.

PURPOSE: Lung cancer is a major cause of cancer-related deaths, emphasizing the importance of early diagnosis. CT-guided percutaneous lung biopsy(CT-PLB) is a valuable method for diagnosing lung lesions, but multiple scans can elevate radiation exposure. This study aims to compare diagnostic efficacy and safety across different CT-PLB protocols. METHODS: 273 consecutive patients who underwent CT-PLB between June 2018 and February 2021 were enrolled, and were divided into standard-dose, conventional low-dose, and experimental low-dose groups. The study mainly evaluated technical success, diagnostic efficacy, radiation dose, complications, and image quality. RESULTS: 93 patients were assigned to standard-dose group, 85 to conventional low-dose group, and 95 to experimental low-dose group. Technical success rates in these groups were 97.9%, 100%, and 97.9%, respectively. Procedure-related complications rates were similar across the groups(pneumothorax:p=0.71, hemorrhage:p=0.59). Sensitivity, specificity, and overall diagnostic accuracy were comparable among three groups(p=0.59,1.0,0.65), with respective values of 90.5%, 100%, and 93.2% in standard-dose group, 88.1%, 100%, and 90.5% in conventional low-dose group, and 91.9%, 100%, and 93.4% in experimental low-dose group. The effective dose (ED) in the experimental low-dose group was significantly lower compared to both the standard-dose and conventional low-dose CT-PLB groups[ED: 1.49(1.0∼1.97) mSv vs 5.42(3.92∼6.91) mSv vs 3.15(2.52∼4.22) mSv, p<0.001]. CONCLUSIONS: This study has developed a standardized six-step procedure for CT-PLB using experimental low-dose settings. It can achieve comparable diagnostic efficacy to conventional low-dose and standard-dose CT-PLB protocols while substantially reducing radiation exposure. These findings indicate that the experimental low-dose protocol could serve as a safe and effective alternative for CT-PLB.

Mapping trade-offs among key ecosystem functions in tidal marsh to inform spatial management policy for exotic Spartina alterniflora.

Invasive Spartina alterniflora has become a global management challenge in coastal wetlands. China has decided to eradicate it completely, but the high costs and its provision of beneficial ecosystem functions (EF, in the form of blue carbon and coastal protection) have raised concerns about its removal. Here, using the Yangtze Estuary as a case study, we explore a reasonable pathway of S. alterniflora management that balanced control of invasive species and EF. We simulated the spatial patterns of two key EF - blue carbon storage and wave attenuation - and identified appropriate zones for eradicating S. alterniflora based on their trade-offs. We observed contrasting patterns along the land-sea gradient for S. alterniflora community, with a decrease in blue carbon storage and an increase in wave attenuation. Notably, pioneer S. alterniflora near the foreshore displayed a high cluster of blue carbon storage (63.61 ± 7.33 Mg C ha-1) and dissipated nearly 70% of wave energy by a width of 163 m. The trade-offs between the two EF indicated that the eradication project should be implemented along the seawall rather than the foreshore. Even in the scenario of prioritized shore defense with the largest eradication zone, S. alterniflora still stored 43.1% more carbon (10.67 Gg C) compared to complete eradication and dissipated over 70% of wave energy in extreme events. Our study innovatively integrates eradication and reservation in S. alterniflora management, providing a sustainable and flexible spatial strategy that meets the needs of stakeholders.

Primary vaginal signet ring cell carcinoma.

Primary vaginal cancer is rare, accounting for only 2% of all gynecological malignant tumors. Primary vaginal cell carcinoma is mainly squamous cell carcinoma, accounting for about 90%, and adenocarcinoma only accounts for 8-10%. Primary signet ring cell carcinoma of vagina is rare and has not been reported in the literature. This paper reports a case of signet ring cell carcinoma in vagina.

A diagnostic scoring model of ENKTCL in the nose-Waldeyer's ring based on logistic regression: Differential diagnosis from DLBCL.

Objective: To establish a logistic regression model based on CT and MRI imaging features and Epstein-Barr (EB) virus nucleic acid to develop a diagnostic score model to differentiate extranodal NK/T nasal type (ENKTCL) from diffuse large B cell lymphoma (DLBCL). Methods: This study population was obtained from two independent hospitals. A total of 89 patients with ENKTCL (n = 36) or DLBCL (n = 53) from January 2013 to May 2021 were analyzed retrospectively as the training cohort, and 61 patients (ENKTCL=27; DLBCL=34) from Jun 2021 to Dec 2022 were enrolled as the validation cohort. All patients underwent CT/MR enhanced examination and EB virus nucleic acid test within 2 weeks before surgery. Clinical features, imaging features and EB virus nucleic acid results were analyzed. Univariate analyses and multivariate logistic regression analyses were performed to identify independent predictors of ENKTCL and establish a predictive model. Independent predictors were weighted with scores based on regression coefficients. A receiver operating characteristic (ROC) curve was created to determine the diagnostic ability of the predictive model and score model. Results: We searched for significant clinical characteristics, imaging characteristics and EB virus nucleic acid and constructed the scoring system via multivariate logistic regression and converted regression coefficients to weighted scores. The independent predictors for ENKTCL diagnosis in multivariate logistic regression analysis, including site of disease (nose), edge of lesion (blurred), T2WI (high signal), gyrus like changes, EB virus nucleic acid (positive), and the weighted score of regression coefficient was 2, 3, 4, 3, 4 points. The ROC curves, AUCs and calibration tests were carried out to evaluate the scoring models in both the training cohort and the validation cohort. The AUC of the scoring model in the training cohort were 0.925 (95% CI, 0.906-0.990) and the cutoff point was 5 points. In the validation cohort, the AUC was 0.959 (95% CI, 0.915-1.000) and the cutoff value was 6 points. Four score ranges were as follows: 0-6 points for very low probability of ENKTCL, 7-9 points for low probability; 10-11 points for middle probability; 12-16 points for very high probability. Conclusion: The diagnostic score model of ENKTCL based on Logistic regression model which combined with imaging features and EB virus nucleic acid. The scoring system was convenient, practical and could significantly improve the diagnostic accuracy of ENKTCL and the differential diagnosis of ENKTCL from DLBCL.

Structural characterization and antinociceptive activity of polysaccharides from Anoectochilus elatus.

Anoectochilus elatus is a new record species from Yunnan province in China discovered by our group in 2018, used in folk as the most popular Anoectochilus species A. roxburghii for medicinal and culinary purposes. The crude polysaccharide of Anoectochilus elatus (AEP) exhibited significant antinociceptive effects against both chemical and thermal nociception in vivo. Bio-guided isolation identified GJXL-1 as the leading analgesic polysaccharide in AEP. Detailed structural analyses rationalized GJXL-1 (molecular weight: 10.3 kDa) as an α-D-1,4-linked glucan unexpectedly branched at O-3, and O-6 position. GJXL-1 dose-dependently suppressed acetic acid-induced writhing in mice and decreased the serum levels of NO, IL-6 and TNF-α, which also repressed the licking times in both the first and second phases in formalin test. Furthermore, only L-nitroarginine partly reversed the analgesic activity of GJXL-1, indicating that GJXL-1's efficacy was partially mediated by NO regulation, possibly through inhibiting IRAK4/TAK1/NF-κB signaling pathway, and modulating gut microbiota and short-chain fatty acids production. In addition, the motor impairment and hypnotic effects of GJXL-1 were excluded. Our study suggests that GJXL-1 can be regarded as a promising and safe drug candidate for diverse pain disorders, and also a promising prebiotic candidate to maintain intestinal homeostasis and promote human gut health.

MRI Extraprostatic Extension Grade: Accuracy and Clinical Incremental Value in the Assessment of Extraprostatic Cancer.

Objective: The purpose was to compare the accuracy of extraprostatic extension (EPE) grade on MRI predicting EPE with Partin tables, Memorial Sloan Kettering Cancer Center nomogram (MSKCCn), and combined models and to analyze the clinical incremental value of EPE grade. Materials and Methods: 105 prostate cancer patients confirmed by pathology after radical prostatectomy in our hospital from 2017 to 2021 were selected. The clinical stage, PSA, Gleason score, number of positive biopsy cores, and percentage of positive biopsy cores were recorded. Evaluate EPE grade according to EPE grade criteria, and calculate the probability of predicting EPE with Partin tables and MSKCCn. EPE grade is combined with Partin tables and MSKCCn to construct EPE grade+Partin tables and EPE grade+MSKCCn models. Calculate the area under the curve (AUC), sensitivity, and specificity of EPE grade, Partin tables, MSKCCn, EPE grade+Partin tables, and EPE grade+MSKCCn and compare their diagnostic efficacy. The clinical decision curve was used to analyze the clinical net income of each prediction scheme. Results: The AUC of EPE grade was 0.79, Partin tables was 0.50, MSKCCn was 0.78, the EPE grade+Partin table model was 0.79, and the EPE grade+MSKCCn model was 0.83. After EPE grade was combined with Partin tables and MSKCCn, the diagnostic efficiency of clinical model was significantly improved (P < 0.05). There was no significant difference in the diagnostic efficacy of the combined model compared with the single EPE grade (P > 0.05). The calibration curve of the combined model shows that it has a good calibration degree for EPE. In the analysis of the decision curve, the net income of the EPE grade is higher than that of Partin tables and MSKCCn and is equal to the EPE grade+Partin tables and is slightly lower than that of EPE grade+MSKCCn. The clinical net income of the combined model is obviously higher than that of individual clinical models. Conclusion: The accuracy of EPE classification in predicting prostate cancer EPE is high, and combined with the clinical model, it can significantly improve the diagnostic efficiency of the clinical model and increase the clinical benefit.

Analysis of serum homocysteine concentration in patients less than 35 years of age with polycystic ovary syndrome and hyperandrogenism.

OBJECTIVES: An increase in homocysteine (Hcy) concentration is closely related to polycystic ovary syndrome (PCOS). This study aimed to further explore serum homocysteine concentration and its influencing factors in clinically young (≤ 35 years) patients with PCOS and hyperandrogenism. MATERIAL AND METHODS: An electrochemical immunoassay was used to investigate the changes in serum homocysteine and related indexes in clinically young patients with PCOS and hyperandrogenism, and the results were statistically analyzed. RESULTS: Serum homocysteine concentration in clinically young patients with PCOS and hyperandrogenism (n = 208) was found to be significantly higher than in the control group (n = 663) (15.21 ± 9.99 vs. 12.56 ± 7.20 µmol/L, p < 0.0001), and the total testosterone concentration (1.65 ± 0.68 ng/mL) was higher in the PCOS group than in the control group (1.52 ± 0.58 ng/mL), p = 0.007. The receiver operating characteristic curve showed that the area under the curve of homocysteine in predicting PCOS was 0.606, and the 95% confidence interval (CI) was 0.563-0.650 (p < 0.001). The homocysteine concentrations of the two groups were graded, and it was found that the percentage of patients with homocysteine levels > 15 µmol/L was 26.92% in the PCOS group and 19.15% in the control group; the difference between the two groups was statistically significant (p = 0.0143). The serum homocysteine levels of the two groups were higher in obese patients than in non-obese patients (normal weight vs. overweight), and the difference in the control group was statistically significant. CONCLUSIONS: Serum homocysteine concentration in clinically young patients with PCOS and hyperandrogenism is elevated, so hyperhomocysteinemia can be used as one of the potential indicators of PCOS. In the process of the diagnosis and treatment of patients with PCOS, serum homocysteine concentration and body weight should both be considered.

Flurochloridone induces responses of free radical reactions and energy metabolism disorders to BRL-3A cell.

Flurochloridone (FLC), a wildly used herbicide, could induce hepatotoxicity after long-term exposure to male rat, in addition to its reactive oxygen species (ROS)-dependent reproductive toxicity. The hepatotoxicity effect and mechanism was investigeted using 1, 10 and 100 µmol L-1 FLC treated BRL-3A liver cell in this study. The function of mitochondrial respiration, glycolysis rate and real time ATP production rate are determined by seahorse XF analyzer, and the bio-transformers of FLC, intermediates of TCA cycle and glycolysis, and related amino acids are determined and identified by [U-13C] Glucose metabolic flux technology based on UPLC-HRMS. The mRNA expression of cytochrome P450s and the key regulatory enzymes of glucose metabolism and γ- glutamyl cycle pathway. The protein expressions of protein kinase B (AKT) and glycogen synthase kinase-3 beta (GSK-3ß) were determined. The results show dechlorination and glutathione (GSH) conjugate products of FLC are predominant bio-transformmers after 24 h treatment in BRL-3A cell. FLC could enhance glycolysis function and inhibit mitochondrial aerobic respiratory, which is accompanied by the decreased total ATP level and ATP produced rate. Increased glucose-6-phosphate, fructose-6-phosphate, pyruvate and lactate levels, and elevated level of GSH and its precursor 5-glutamate-cysteine (γ-Glu-Cys) are observed in FLC treated cells, which indicates that energy metabolism dysfunction and GSH accumulation could be potentially mediated by activating γ- Glutamyl cycle pathway. Conclusively, FLC induced hepatotoxicity could be potentially related to some free radical reactions, including inhibiting mitochondrial function, glucose metabolism via glycolysis, regulating γ- glutamyl cycle pathway to promote reactive oxygen species (ROS) level, and then induced cell apoptosis by inhibiting AKT/GSK-3ß signal.

Profiling of pesticides and pesticide transformation products in Chinese herbal teas.

Herbal teas have potential health benefits, but they also contain a variety of pesticides and pesticide transformation products (PTPs) that might brings health risks. Our study maps the pesticides and PTPs in two herbal teas (chrysanthemum and Lusterleaf Holly) from two main producing areas in China. Almost all 122 samples contain pesticides, with concentration ranging from 0.0005 to 10.305 mg/kg. Nearly 40% carbendazim and imidacloprid in chrysanthemum teas and λ-cyhalothrin in Lusterleaf Holly have higher concentration levels than the values permitted in EC Regulation No. 396/2005. Distinct distributions of pesticides were found in different teas and production areas. Eight PTPs were identified along with their parents, and were confirmed using a biosynthetic strategy. Acute, chronic and cumulative health risk assessments of pesticides revealed acceptable results. Our study uncovers the profile of pesticides in herbal teas, and provides new insight into discovering the potential environmental pollution and food contaminants.

Intestinal farnesoid X receptor signaling controls hepatic fatty acid oxidation.

In addition to maintaining bile acid, cholesterol and glucose homeostasis, farnesoid X receptor (FXR) also regulates fatty acid ß-oxidation (FAO). To explore the different roles of hepatic and intestinal FXR in liver FAO, FAO-associated metabolites, including acylcarnitines and fatty acids, and FXR target gene mRNAs were profiled using an integrated metabolomic and transcriptomic analysis in control (Fxrfl/fl), liver-specific Fxr-null (FxrΔHep) and intestine-specific Fxr-null (FxrΔIE) mice, treated either with the FXR agonist obeticholic acid (OCA) or vehicle (VEH). Activation of FXR by OCA treatment significantly increased fatty acyl-CoA hydrolysis (Acot1) and decreased FAO-associated mRNAs in Fxrfl/fl mice, resulting in reduced levels of total acylcarnitines and relative accumulation of long/medium chain acylcarnitines and fatty acids in liver. FxrΔHep mice responded to OCA treatment in a manner similar to Fxrfl/fl mice while FxrΔIE mice responded differently, thus illustrating that intestinal FXR plays a critical role in the regulation of hepatic FAO. A significant negative-correlation between intestinal FXR-FGF15 and hepatic CREB-PGC1A pathways was observed after both VEH and OCA treatment, suggesting that OCA-induced activation of the intestinal FXR-FGF15 axis downregulates hepatic PGC1α signaling via inactivation of hepatic CREB, thus repressing FAO. This mechanism was confirmed in experiments based on human recombinant FGF19 treatment and intestinal Fgf15-null mice. This study revealed an important role for the intestinal FXR-FGF15 pathway in hepatic FAO repression.

Investigating the Effects of Dapagliflozin on Cardiac Function, Inflammatory Response, and Cardiovascular Outcome in Patients with STEMI Complicated with T2DM after PCI.

OBJECTIVE: To explore the effect of dapagliflozin on cardiac function, inflammation, and cardiovascular outcome in patients with ST-segment elevation myocardial infarction (STEMI) combined with type 2 diabetes (T2DM) after percutaneous coronary intervention (PCI). METHODS: 70 patients with STEMI and T2DM were divided into the control group (n = 35) and the observation group (n = 35). Before surgery, patients in both groups were given conventional treatments such as coronary expansion, antiplatelet, anticoagulation, and thrombolysis, and PCI was performed. After the operation, both groups were given conventional antiplatelet, anticoagulation, lipid-lowering, and hypoglycemic treatments. On this basis, the observation group was treated with dapagliflozin tablets for 24 weeks. We observe and compare the left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD) and left ventricular ejection fraction (LVEF), myocardial enzyme spectrum, inflammatory reaction, and occurrence of adverse cardiovascular events (MACE) of the two groups of patients before and after treatment. RESULTS: After treatment, the LVEDD and LVESD of the two groups were lower than those before treatment, and the observation group was lower than the control group (P < 0.05). The LVEF of both groups was higher than that before treatment, and the observation group was higher than the control group (P < 0.05). After treatment, the levels of two groups' patients' creatine kinase (CK), creatine kinase isoenzyme (CK-MB), and troponin I (cTnI) were all lower than those before treatment, and the observation group patients were all lower than the control group (P < 0.05). After treatment, the levels of serum myeloperoxidase (MPO), C-reactive protein (CRP), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) in the two groups were all lower than those before treatment, and the observation group patients were all lower than the control group (P < 0.05). After treatment, there was no statistical difference between the two groups of patients in cardiogenic death, recurrent myocardial infarction, and other adverse cardiovascular events (P > 0.05). But, the incidence of severe arrhythmia and heart failure in the observation group were both lower than those in the control group (P < 0.05). Kaplan-Meier survival curve analysis showed that the median survival time without MACE in the observation group was higher than that in the control group (P < 0.05). CONCLUSION: Dapagliflozin treatment for patients with STEMI combined with T2DM after PCI can improve cardiac function to certain extent, reduce inflammation, and will reduce the incidence of adverse cardiovascular outcomes.

Novel Strategy for Mining and Identification of Acylcarnitines Using Data-Independent-Acquisition-Based Retention Time Prediction Modeling and Pseudo-Characteristic Fragmentation Ion Matching.

It is a challenging work to screen, identify, and quantify acylcarnitines in complex biological samples. A method, based on the retention time (RT) prediction and data-independent acquisition strategies, was proposed for the large-scale identification of acylcarnitines using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS). Relative cumulative eluotropic strength was introduced as a novel descriptor in building a linear prediction model, which not only solves the problem that acylcarnitines with long carbon chains cannot be well predicted in traditional models but also proves its robustness and transferability across instruments in two data sets that were acquired in distinct chromatography conditions. The accessibility of both predictive RT and MS2 spectra of suspect features effectively reduced about 30% false-positive results, and consequently, 150 and 186 acylcarnitines were identified in the rat liver and human plasma (NIST SRM 1950), respectively. This method provides a new approach in large-scale analysis of acylcarnitine in lipidomic studies and can also be extended to the analysis of other lipids.

Oxytocin weakens self-other distinction in males during empathic responses to sadness: an event-related potentials study.

By making use of event-related potential (ERP) technology, a randomized, double-blind, between-subject design study was performed in order to investigate whether OXT can weaken men's self-other distinction during empathic responses to sad expressions. In the two experimental tasks, 39 male subjects were asked to either evaluate the emotional state shown in a facial stimulus (other-task) or to evaluate their own emotional responses (self-task). The results revealed that OXT reduced the differences in P2 (150-200 ms) amplitudes between sad and neutral expressions in the self-task but enhanced P2 to sad expressions in the other-task, indicating OXT's role in integrating the self with others instead of separating them. In addition, OXT also reduced the LPC (400-600 ms) amplitudes between sad-neutral expressions in the self-task, implying that OXT's weakening effects on the self-other distinction could occur at both the early and late cognitive control stages of the empathic response.

A systemic workflow for profiling metabolome and lipidome in tissue.

Simple metabolome and lipidome sample preparation procedures involving two successive extractions using small pieces of tissue, and a subsequent metabolite identification (MetID) strategy were developed. The sample preparation can significantly circumvent incomplete analysis due to insufficient amounts of tissue as a result of splitting into several aliquots for multiple measurements, with advantages over the similar previously reported methods in metabolite coverage, extraction efficiency, method robustness and friendly experimental operation. A MetID strategy, based on the integration of MS information mining (including adduct ions, in-source CID, MS information from both ESI (+) and ESI (-), characteristic fragmentation ions (CFIs), constant neutral losses (CNLs) and multimers) and in silico MS simulation, was demonstrated. A large number of adduct ions (83 features), in-source CID (123 features), ESI (+/-) ionization (20 features), CFIs& CNLs (more than 120 features) and multimers (17 features) were mined by manually or in silico recognition/filtering, which provide the most suspicious structures for subsequent in silico MS simulation. The unknown features presented the same score distribution as the known (83 features) features with scores ≥25% (geomean score: 52%) and with satisfactory match for the main ions of interest. The MS/MS noise and fragment ions of coeluted quasi-molecular ions of interest are the main reason for the low score in the simulation. Manual check/evaluation is always suggested for the simulation with a score less than 50%. This strategy presents satisfactory performance with 2.5 times more metabolites structurally characterized compared with that of the traditional method based on accurate-mass-based MS and MS/MS library matching. This strategy would be useful for potentially identifying metabolites without available MS/MS information in the library.

Accuracy of Multi-Slice Spiral Computed Tomography for Preoperative Tumor Node Metastasis (TNM) Staging of Colorectal Carcinoma.

BACKGROUND With the advances in imaging technologies, multi-slice spiral computed tomography (MSCT) has demonstrated superiority in the diagnosis and staging of colorectal carcinoma. In the current study, preoperative TNM staging of colorectal carcinoma by using MSCT was conducted and compared with the corresponding postoperative pathological examination findings, in order to evaluate the accuracy of preoperative MSCT for TNM staging. MATERIAL AND METHODS Combinations of biphasic or triphasic enhanced-phase MSCT scans were obtained for 76 patients with colorectal carcinoma, and the TNM stage was determined based on imaging reconstruction from various angles and perspectives to display the size, location, and affected range of tumors. The preoperative TNM stage was compared with the postoperative pathological stage, and the consistency between the 2 methods was tested by the k test using SPSS 17.0 software. RESULTS Among the different combinations of enhanced-phase MSCT scanning, triphasic MSCT imaging, comprising the arterial, portal venous, and delayed phases, showed the highest accuracy rates, at 81.6% (62/76), 82.89% (63/76), and 96.1% (73/76) for T, N, and M staging, respectively, with k values of 0.72, 0.65, and 0.56, respectively, indicating consistency with the postoperative pathological staging. CONCLUSIONS Combined MSCT scanning comprising the arterial phase, portal venous phase, and delayed phase showed satisfying consistency with the postoperative pathological analysis results for TNM staging of colorectal carcinoma. Thus, MSCT is an important clinical value for improving the accuracy of TNM staging and for planning the appropriate colorectal cancer treatment.

An Ectosteric Inhibitor of Cathepsin K Inhibits Bone Resorption in Ovariectomized Mice.

The potent cathepsin K (CatK) inhibitor, Tanshinone IIA sulfonic sodium (T06), was tested for its in vitro and in vivo antiresorptive activities. T06 binds in an ectosteric site of CatK remote from its active site and selectively inhibits collagen degradation with an IC50 value of 2.7 ± 0.2 µM (CatK:T06 molar ratio of 1:5). However, it does not suppress fluorogenic peptide cleavage and gelatinolysis at a 2500-fold molar excess. Contrary to active site-directed CatK inhibitors, such as odanacatib, T06 suppresses bone resorption in both human and mouse osteoclasts equally well (IC50 value for human and mouse osteoclasts: 237 ± 60 nM and 245 ± 55 nM, respectively) and its antiresorptive activity is fully reversible in both cell types. Moreover, T06 affects neither the metabolic activity of osteoclasts nor osteoclastogenesis. In in vivo studies, 40 mg T06/kg/d given to 12-week-old ovariectomized (OVX) mice for 3 months reduced plasma CTx-1 by 20% and increased osteoblast numbers and plasma P1NP by ∼28% when compared with the OVX control. µCT analysis of T06-treated OVX mice showed a 35% increase in bone mineral density and other femoral trabecular bone parameters when compared with OVX animals. T06 did not alter the number of osteoclasts, had no estrogenic effect on the uterus, did not change plasma estradiol levels, and did not inhibit fibroblast-mediated TGF-ß1 processing or degradation and cognitive functions in OVX mice. This study indicates that the ectosteric inhibitor, T06, is a selective antiresorptive CatK inhibitor that may overcome the shortcomings of side effect-prone active site-directed drugs, which all failed in clinical trials. © 2017 American Society for Bone and Mineral Research.

High Expression of Human Cathepsin S by Recombinant Pichia pastoris with Cod Skin as an Organic Co-Nitrogen Source.

Human cathepsin S production by recombinant Pichia pastoris using cod skin as the co-nitrogen source was investigated in this study. The addition of carbon sources of glycerol in the fed-batch phase and of methanol in the induction stage was also investigated. A new approach to the highly expression of human cathepsin S was developed using 90 g/L of cod skin (wet weight). After 24 h of the initial fermentation, 4% glycerol (v/v, glycerol/culture) was added once to enhance the cell density (OD600) in the cultivation. Then, adding and maintaining methanol at 0.5% (v/v, methanol/cultivation) after about 48 h of fermentation achieved a high expression of human cathepsin S in a 5-L bioreactor. The results demonstrate that the maximum activity of human cathepsin S in the fermentation supernatant reached 7,152 U/L after 96 h of methanol induction. The methylotrophic yeast P. pastoris grown in the medium containing cod skin (90 g/L) as the co-nitrogen source provided a 21% higher cell density (OD600) and 18.3% higher human cathepsin S yield than P. pastoris grown in BMGY medium. For the first time, human cathepsin S was successfully expressed by P. pastoris with cod skin as the co-nitrogen source. The glycerol fed-batch controlling strategy and method of maintaining methanol at a constant concentration of 0.5% (v/v, methanol/cultivation) in the induction stage was efficient for P. pastoris growth and the expression of human cathepsin S.

Comparative proteomic and metabolomic analysis reveal the antiosteoporotic molecular mechanism of icariin from Epimedium brevicornu maxim.

ETHNOPHARMACOLOGICAL RELEVANCE: Icariin, a principal flavonoid glycoside of Epimedium brevicornu Maxim, has been widely proved to possess antiosteoporotic activity with promoting bone formation and decreasing bone resorption. However, the involving mechanisms remain unclear. AIM OF THE STUDY: To clear a global insight of signal pathways involved in anti-osteoporotic mechanism of icariin at proteins and metabolites level by integrating the proteomics and NMR metabonomics, in a systems biology approach. MATERIAL AND METHODS: Mice were divided into sham, OVX model and icariin-treated OVX group, after 90 days treatment, difference gel electrophoresis combined with MALDI-TOF/TOF proteomics analysis on bone femur and serum metabolomics were carried out for monitor intracellular processes and elucidate anti-osteoporotic mechanism of icariin. Osteoblast and osteoclast were applied to evaluate the potential signal pathways. RESULTS: Twenty three proteins in bone femur, and 8 metabolites in serum, were significantly altered and identified, involving in bone remodeling, energy metabolism, cytoskeleton, lipid metabolism, MAPK signaling, Ca2+ signaling et, al. Furthermore, animal experiment show icariin could enhance the BMD and BMC, decrease CTX-I level in ovariectomized mice. The mitochondrial membrane potential and the intracellular ATP levels were increased significantly, and the cytoskeleton were improved in icariin-treatment osteoblast and osteoclast. Icariin also increased mRNA expression of Runx2 and osterix of OB, decreased CTR and CAII mRNA expression and protein expression of P38 and JNK. However, icariin did not reveal any inhibition of the collagenolytic activity of cathepsin K, mRNA expression of MMP-9 and protein expression of ERK in osteoclast. CONCLUSION: we consider icariin as multi-targeting compounds for treating with osteoporosis, involve initiating osteoblastogenesis, inhibiting adipogenesis, and preventing osteoclast differentiation.