Neuropeptides and hormones in hypothalamus-pituitary axis of Chinese sturgeon (Acipenser sinensis).

The hypothalamus and pituitary serve as important neuroendocrine center, which is able to secrete a variety of neuropeptides and hormones to participate in the regulation of reproduction, growth, stress and feeding in fish. Chinese sturgeon is a basal vertebrate lineage fish with a special evolutionary status, but the information on its neuroendocrine system is relatively scarce. Using the transcriptome data on the hypothalamus-pituitary axis of Chinese sturgeon as reference, we found out 46 hypothalamus neuropeptide genes, which were involved in regulation of reproduction, growth, stress and feeding. The results of sequence alignment showed that the neuroendocrine system of Chinese sturgeon evolves slowly, which confirms that Chinese sturgeon is a species with a slow phenotypic evolution rate. In addition, we also isolated six pituitary hormones genes from Chinese sturgeon, including reproductive hormones: follicle-stimulating homone (FSH) and luteinizing hormone (LH), growth-related hormones: growth hormone (GH)/prolactin (PRL)/somatolactin (SL), and stress-related hormone gene: proopiomelanocortin (POMC). Similar to teleost, immunostaining localization analysis in Chinese sturgeon pituitary showed that LH and FSH were located in the pituitary proximal pars distalis, SL was located in the pituitary rostral pars distalis, and POMC was located in the pituitary pars intermedia and pituitary rostral pars distalis. This study will give a contribution to enrich our information on the neuroendocrine system in Chinese sturgeon.

Temperature dependence of dielectric properties of blood at 10 Hz-100 MHz.

The temperature dependence of the dielectric properties of blood is important for studying the biological effects of electromagnetic fields, electromagnetic protection, disease diagnosis, and treatment. However, owing to the limitations of measurement methods, there are still some uncertainties regarding the temperature characteristics of the dielectric properties of blood at low and medium frequencies. In this study, we designed a composite impedance measurement box with high heat transfer efficiency that allowed for a four/two-electrode measurement method. Four-electrode measurements were carried out at 10 Hz-1 MHz to overcome the influence of electrode polarization, and two-electrode measurements were carried out at 100 Hz-100 MHz to avoid the influence of distribution parameters, and the data was integrated to achieve dielectric measurements at 10 Hz-100 MHz. At the same time, the temperature of fresh blood from rabbits was controlled at 17-39°C in combination with a temperature-controlled water sink. The results showed that the temperature coefficient for the real part of the resistivity of blood remained constant from 10 Hz to 100 kHz (-2.42%/°C) and then gradually decreased to -0.26%/°C. The temperature coefficient of the imaginary part was positive and bimodal from 6.31 kHz to 100 MHz, with peaks of 5.22%/°C and 4.14%/°C at 126 kHz and 39.8 MHz, respectively. Finally, a third-order function model was developed to describe the dielectric spectra at these temperatures, in which the resistivity parameter in each dispersion zone decreased linearly with temperature and each characteristic frequency increased linearly with temperature. The model could estimate the dielectric properties at any frequency and temperature in this range, and the maximum error was less than 1.39%, thus laying the foundation for subsequent studies.

Reproductive Regulation of PrRPs in Teleost: The Link Between Feeding and Reproduction.

Prolactin-releasing peptide (PrRP), a sort of vital hypothalamic neuropeptide, has been found to exert an enormous function on the food intake of mammals. However, little is known about the functional role of PrRP in teleost. In the present study, two PrRP isoforms and four PrRP receptors were isolated from grass carp. Ligand-receptor selectivity displayed that PrRP1 preferentially binds with PrRP-R1a and PrRP-R1b, while PrRP-R2a and PrRP-R2b were special receptors for PrRP2. Tissue distribution indicated that both PrRPs and PrRP-Rs were highly expressed in the hypothalamus-pituitary-gonad axis and intestine, suggesting a latent function on food intake and reproduction. Using grass carp as a model, we found that food intake could significantly induce hypothalamus PrRP mRNA expression, which suggested that PrRP should be also an anorexigenic peptide in teleost. Interestingly, intraperitoneal (IP) injection of PrRPs could significantly induce serum luteinizing hormone (LH) secretion and pituitary LHß and GtHα mRNA expression in grass carp. Moreover, using primary culture grass carp pituitary cells as a model, we further found that PrRPs could directly induce pituitary LH secretion and synthesis mediated by AC/PKA, PLC/IP3/PKC, and Ca2+/CaM/CaMK-II pathways. Finally, estrogen treatment of prepubertal fish elicited increases in PrRPs and PrPR receptors expression in primary cultured grass carp hypothalamus cells, which further confirmed that the PrRP/PrRPR system may participate in the neuroendocrine control of fish reproduction. These results, taken together, suggest that PrRPs might act as a coupling factor in feeding metabolism and reproductive activities in teleost.

Sintilimab plus a bevacizumab biosimilar (IBI305) versus sorafenib in unresectable hepatocellular carcinoma (ORIENT-32): a randomised, open-label, phase 2-3 study.

BACKGROUND: China has a high burden of hepatocellular carcinoma, and hepatitis B virus (HBV) infection is the main causative factor. Patients with hepatocellular carcinoma have a poor prognosis and a substantial unmet clinical need. The phase 2-3 ORIENT-32 study aimed to assess sintilimab (a PD-1 inhibitor) plus IBI305, a bevacizumab biosimilar, versus sorafenib as a first-line treatment for unresectable HBV-associated hepatocellular carcinoma. METHODS: This randomised, open-label, phase 2-3 study was done at 50 clinical sites in China. Patients aged 18 years or older with histologically or cytologically diagnosed or clinically confirmed unresectable or metastatic hepatocellular carcinoma, no previous systemic treatment, and a baseline Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 were eligible for inclusion. In the phase 2 part of the study, patients received intravenous sintilimab (200 mg every 3 weeks) plus intravenous IBI305 (15 mg/kg every 3 weeks). In the phase 3 part, patients were randomly assigned (2:1) to receive either sintilimab plus IBI305 (sintilimab-bevacizumab biosimilar group) or sorafenib (400 mg orally twice daily; sorafenib group), until disease progression or unacceptable toxicity. Randomisation was done using permuted block randomisation, with a block size of six, via an interactive web response system, and stratified by macrovascular invasion or extrahepatic metastasis, baseline α-fetoprotein, and ECOG performance status. The primary endpoint of the phase 2 part of the study was safety, assessed in all patients who received at least one dose of study drug. The co-primary endpoints of the phase 3 part of the study were overall survival and independent radiological review committee (IRRC)-assessed progression-free survival according to Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 in the intention-to-treat population. The study is registered with ClinicalTrials.gov, NCT03794440. The study is closed to new participants and follow-up is ongoing for long-term outcomes. FINDINGS: Between Feb 11, 2019 and Jan 15, 2020, we enrolled 595 patients: 24 were enrolled directly into the phase 2 safety run-in and 571 were randomly assigned to sintilimab-bevacizumab biosimilar (n=380) or sorafenib (n=191). In the phase 2 part of the trial, 24 patients received at least one dose of the study drug, with an objective response rate of 25·0% (95% CI 9·8-46·7). Based on the preliminary safety and activity data of the phase 2 part, in which grade 3 or worse treatment-related adverse events occurred in seven (29%) of 24 patients, the randomised phase 3 part was started. At data cutoff (Aug 15, 2020), the median follow-up was 10·0 months (IQR 8·5-11·7) in the sintilimab-bevacizumab biosimilar group and 10·0 months (8·4-11·7) in the sorafenib group. Patients in the sintilimab-bevacizumab biosimilar group had a significantly longer IRRC-assessed median progression-free survival (4·6 months [95% CI 4·1-5·7]) than did patients in the sorafenib group (2·8 months [2·7-3·2]; stratified hazard ratio [HR] 0·56, 95% CI 0·46-0·70; p<0·0001). In the first interim analysis of overall survival, sintilimab-bevacizumab biosimilar showed a significantly longer overall survival than did sorafenib (median not reached [95% CI not reached-not reached] vs 10·4 months [8·5-not reached]; HR 0·57, 95% CI 0·43-0·75; p<0·0001). The most common grade 3-4 treatment-emergent adverse events were hypertension (55 [14%] of 380 patients in the sintilimab-bevacizumab biosimilar group vs 11 [6%] of 185 patients in the sorafenib group) and palmar-plantar erythrodysaesthesia syndrome (none vs 22 [12%]). 123 (32%) patients in the sintilimab-bevacizumab biosimilar group and 36 (19%) patients in the sorafenib group had serious adverse events. Treatment-related adverse events that led to death occurred in six (2%) patients in the sintilimab-bevacizumab biosimilar group (one patient with abnormal liver function, one patient with both hepatic failure and gastrointestinal haemorrhage, one patient with interstitial lung disease, one patient with both hepatic faliure and hyperkalemia, one patient with upper gastrointestinal haemorrhage, and one patient with intestinal volvulus) and two (1%) patients in the sorafenib group (one patient with gastrointestinal haemorrhage and one patient with death of unknown cause). INTERPRETATION: Sintilimab plus IBI305 showed a significant overall survival and progression-free survival benefit versus sorafenib in the first-line setting for Chinese patients with unresectable, HBV-associated hepatocellular carcinoma, with an acceptable safety profile. This combination regimen could provide a novel treatment option for such patients. FUNDING: Innovent Biologics. TRANSLATION: For the Chinese translation of the abstract see Supplementary Materials section.

Dielectric Properties of Human Active Liver, Kidney and Spleen Compared to Those of Respective Inactive Tissues, Porcine Tissues and the Data Provided by a Database in the Frequency Range of 10 Hz to 100 MHz.

OBJECTIVE: The purpose of this work is to study whether the active state and species of biological tissues can influence changes in their dielectric properties. METHODS: In this paper, the dielectric properties of liver, kidney and spleen tissues from human active, human inactive and animal tissues are measured in the frequency range of 10 Hz to 100 MHz. The four- and two-electrode methods are used to measure dielectric properties at different frequencies. Statistical analysis and the pattern recognition method are used to compare the dielectric properties of human active tissues, human inactive tissues, animal tissues and data provided by the IFAC database. RESULTS: The results show that the dielectric properties of human active tissues are significantly different from those of human inactive tissues and animal tissues, resulting in a great difference between the dielectric properties provided by the IFAC database and those of human active tissues. The dielectric properties of human active tissues can be identified by the pattern recognition method based on principal component analysis, which further proves that the dielectric properties of human active tissues cannot be replaced. CONCLUSION: The dielectric properties of biological tissues are closely related to the activity and species of tissues. The dielectric properties of human active tissues cannot be replaced by those of human cadaver tissues or animal tissues. SIGNIFICANCE: The significance of this study is suggesting that the IFAC database should be updated with the dielectric properties of human active tissues to provide accurate data for bioelectromagnetics research.

Engineering natural matrices with black phosphorus nanosheets to generate multi-functional therapeutic nanocomposite hydrogels.

Natural polysaccharides and proteins have been widely explored for the preparation of hydrogel matrices due to their promising biocompatibility and biodegradability. However, it is challenging to achieve multiple functions of the hydrophilic matrix through convenient functionalization strategies. Herein we report the facile engineering of a natural matrix with black phosphorus (BP) nanosheets as building blocks to generate a therapeutic nanocomposite hydrogel (BP/Gel) with an array of promising features. BP nanosheets could reinforce the crosslinking networks and significantly promote their capabilities of mineralization. The BP/Gel nanocomposite hydrogel exhibits excellent near infrared (NIR) photothermal performance and good biocompatibility in vitro and in vivo. Upon NIR irradiation, the nanocomposite hydrogel demonstrates efficient photothermal antibacterial features. More remarkably, the BP nanosheet engineered hydrogel matrix is capable of promoting in vitro osteogenesis in the absence of osteoinductive factors, and in the meantime demonstrates significant newborn cranial bone tissue formation in a Sprague-Dawley rat model. These results demonstrate that BP nanosheets could endow the natural matrix with multiple functions including reinforced networks, photothermal performance, enhanced mineralization and bone regeneration, which provides a facile and highly efficient therapeutic strategy for bone tissue engineering.

TCM matrine inducescell arrest and apoptosis with recovery expression of the hepato-specific miR122a in human hepatocellular carcinomaHep G2cell line.

Hepatocellular carcinoma (HCC) accounts for 80% to 90% of liver cancers and it is one of the most prevalent carcinomas throughout the world. Traditional chemotherapy is often developed chemoresistance HCC patients.Matrine is an active component oftraditional Chinese medicine (TCM) and is a promising alternative HCC drug. In this study, the therapeutic effects and the underlying molecular mechanisms of matrine on the human HCC cell lineHep G2 were investigated. High dosage of matrine (1.0 mg/mL) could significantly (P < 0.05) inhibit cell proliferation by 48.39 ± 3.32%, under which cell shrinkage and disruption were observed. Flow cytometry assay showed that the proportion of G1/G0 cells significantly increased, while that of S and G2/M cells significantly decreased after treatment of matrinefor 48 h. These results indicated that cell arrest by matrine appeared. Up-regulation of the hepato-specific miR122a followed by down expression of its targetcyclin G1 (CG1) gene by low concentration of matrine (0.2 mg/mL) was detected using was observed using quantitative real-time PCR, immunohistochemistry (IHC) and western blot assays. In conclusion, matrineinducescell arrest and apoptosis with recovery expression of the hepato-specific miR122a in human hepatocellular carcinoma Hep G2 cell line.

Lipiodol: a potential direct surrogate for cone-beam computed tomography image guidance in radiotherapy of liver tumor.

PURPOSE: To investigate the feasibility of using lipiodol as a direct surrogate for target localization using cone-beam CT (CBCT) image guidance in radiotherapy (RT) of patients with unresectable liver tumors after transarterial chemoembolization. METHODS AND MATERIALS: Forty-six patients with an unresectable solitary liver tumor were enrolled for RT using active breathing control (ABC) and CBCT image guidance after transarterial chemoembolization. Each patient had pre- and posttreatment CBCT in the first 10 fractions of treatment. Lipiodol retention was evaluated using daily CBCT scans, and volume of lipiodol retention in the liver was calculated and compared between planning CT and post-RT CT. Influence of lipiodol on dosimetry was evaluated by measuring doses using an ion chamber with and without the presence of lipiodol. Margin analysis was performed on the basis of both inter- and intrafractional target localization errors. RESULTS: Twenty-eight patients successfully completed the study. The shape and size of lipiodol retention did not vary substantially during the course of treatment. The mean Dice similarity coefficient for the lipiodol volume in pretreatment CT and that in posttreatment CT was 0.836 (range, 0.817-0.885). The maximum change (ratio of the lipiodol volume in pretreatment CT to that in posttreatment CT) was 1.045. The mean dose changes with the presence of <10 mL lipiodol were -1.44% and 0.13% for 6 MV and 15 MV, respectively. With ABC and online CBCT image guidance, clinical target volume-planning target volume margins were determined to be 2.5 mm in the mediolateral direction, 2.9 mm in the anteroposterior direction, and 4.0 mm in the craniocaudal direction. CONCLUSIONS: Lipiodol could be used as a direct surrogate for CBCT image guidance to improve the localization accuracy for RT of liver tumors. Combination of ABC and CBCT image guidance with lipiodol can potentially reduce the clinical target volume-planning target volume margin.

A novel hydrophilic poly(lactide-co-glycolide)/lecithin hybrid microspheres sintered scaffold for bone repair.

Novel 3-D porous scaffolds made of sintered poly(lacide-co-glycolide) (PLGA)/lecithin hybrid microspheres (PLGA/Lec-SMS) were developed and investigated. The addition of lecithin in PLGA bulk successfully managed the desired hydrophilic modification without sacrificing bulk properties. The outcomes were verified with infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and contact angle analyses. Specifically, this model of scaffold gained significant improvement in mechanical (mainly compressive) strength upon an optimization of lecithin fractions aligning with sintering conditions. Given a perspective of bone tissue engineering use, human fetal osteoblasts were seeded into a series of these PLGA/Lec-SMS scaffolds upon which key parameters of cytocompatibility and osteoconductivity (including cell viability, alkaline phosphatase activity, calcium secretion, and osteogenic genes expression) were assessed. Osteoblasts seeded on PLGA scaffolds with 5 wt % lecithin demonstrated high cell viability and alkaline phosphatase activity. Moreover, elevated lecithin also enhanced the expression of type I collagen. Taken together, these results suggest PLGA/Lec-SMS are promising scaffolds for bone repair.

A protein/antibiotic releasing poly(lactic-co-glycolic acid)/lecithin scaffold for bone repair applications.

Novel poly(lactic-co-glycolic acid) (PLGA)-hybridizing-lecithin scaffolds loaded with drug or protein were prepared with water/oil/water techniques and sintering microspheres technique. In such fabricated composite scaffolds (abbreviated "PLGA/Lec-SMS"), the introduction of lecithin component has been proven capable of largely enhancing Gentamicin (GS) and protein (Bovine Serum Albumin) encapsulation efficiency. The in vitro GS and BSA releasing profiles of PLGA/Lec-SMS system were plotted basing over 60 days' and 18 days' data collection, respectively. It indicates a sustained releasing tendency despite a burst at the very beginning. The antibacterial properties of GS-laden scaffolds were determined in vitro, and the antibacterial activity of scaffolds was enhanced by incorporating lecithin into PLGA bulks. Additionally, mesenchymal stem cells (MSCs) were seeded onto PLGA-SMS and PLGA/Lec-SMS in vitro. The outcome confirmed PLGA/Lec(5%)-SMS functions to improve MSC proliferation and also to enhance general ALP production and calcium secretion which is the vital markers for osteogenesis. In conclusion, this newly designed antibiotic releasing PLGA/Lec-SMS is promising for bone-repairing therapeutics.