Directed osteogenic differentiation of human bone marrow mesenchymal stem cells via sustained release of BMP4 from PBVHx-based nanoparticles.

Bone Morphogenetic Protein 4 (BMP4) is crucial for bone and cartilage tissue regeneration, essential in medical tissue engineering, cosmetology, and aerospace. However, its cost and degradation susceptibility pose significant clinical challenges. To enhance its osteogenic activity while reducing dosage and administration frequency, we developed a novel long-acting BMP4 delivery system using poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) (PBVHx) nanoparticles with soybean lecithin-modified BMP4 (sBP-NPs). These nanoparticles promote directed osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) through sustained BMP4 release. sBP-NPs exhibited uniform size (100-200 nm) and surface charges, with higher BMP4 entrapment efficiency (82.63 %) compared to controls. After an initial burst release within 24 h, sBP-NPs achieved 80 % cumulative BMP4 release within 20 days, maintaining levels better than control BP-NPs with unmodified BMP4. Co-incubation and nanoparticle uptake experiments confirmed excellent biocompatibility of sBP-NPs, promoting hBMSC differentiation towards osteogenic lineage with increased expression of type I collagen, calcium deposition, and ALP activity (> 20,000 U/g protein) compared to controls. Moreover, hBMSCs treated with sBP-NPs exhibited heightened expression of osteogenic genetic markers, surpassing control groups. Hence, this innovative strategy of sustained BMP4 release from sBP-NPs holds potential to revolutionize bone regeneration in minimally invasive surgery, medical cosmetology or space environments.

Combination Therapies to Improve the Efficacy of Immunotherapy in Triple-negative Breast Cancer.

Immune checkpoint inhibition combined with chemotherapy is currently approved as first-line treatment for patients with advanced PD-L1-positive triple-negative breast cancer (TNBC). However, a significant proportion of metastatic TNBC is PD-L1-negative and, in this population, chemotherapy alone largely remains the standard-of-care and novel therapeutic strategies are needed to improve clinical outcomes. Here, we describe a triple combination of anti-PD-L1 immune checkpoint blockade, epigenetic modulation thorough bromodomain and extra-terminal (BET) bromodomain inhibition (BBDI), and chemotherapy with paclitaxel that effectively inhibits both primary and metastatic tumor growth in two different syngeneic murine models of TNBC. Detailed cellular and molecular profiling of tumors from single and combination treatment arms revealed increased T- and B-cell infiltration and macrophage reprogramming from MHCIIlow to a MHCIIhigh phenotype in mice treated with triple combination. Triple combination also had a major impact on gene expression and chromatin profiles shifting cells to a more immunogenic and senescent state. Our results provide strong preclinical evidence to justify clinical testing of BBDI, paclitaxel, and immune checkpoint blockade combination.

Arterial blood gas analysis aids early differential diagnosis and treatment of primary and secondary hypokalaemic periodic paralysis.

This study aimed to examine changes in electrolytes and acid-base status in primary and secondary hypokalaemic periodic paralysis (HypoPP), which will help early differential diagnosis of HypoPP. A total of 64 HypoPP patients were enrolled and relevant data from clinical records was collected. Overall, 64 patients (mean age 28.2±7.3 years) of which 58(91%) were males, with 39, 11 and 14 patients, respectively, diagnosed as primary HypoPP, thyrotoxic HypoPP, and other secondary HypoPPs at discharge, were assessed. Those with HypoPP secondary to conditions other than hyperthyroidism were more likely to develop acid-base imbalance (p<0.001); they had higher pH (p=0.046) and HCO3 levels (p=0.014) at baseline, and needed a higher dose of potassium supplement before the serum potassium level returned to normal (p=0.007) and a longer time to regain full muscle strength (p=0.004), compared with those with primary or thyrotoxic HypoPP. Emergent arterial blood gas analysis may aid early differential diagnosis of patients with primary and secondary HypoPP.

Dissecting spatial heterogeneity and the immune-evasion mechanism of CTCs by single-cell RNA-seq in hepatocellular carcinoma.

Little is known about the transcriptomic plasticity and adaptive mechanisms of circulating tumor cells (CTCs) during hematogeneous dissemination. Here we interrogate the transcriptome of 113 single CTCs from 4 different vascular sites, including hepatic vein (HV), peripheral artery (PA), peripheral vein (PV) and portal vein (PoV) using single-cell full-length RNA sequencing in hepatocellular carcinoma (HCC) patients. We reveal that the transcriptional dynamics of CTCs were associated with stress response, cell cycle and immune-evasion signaling during hematogeneous transportation. Besides, we identify chemokine CCL5 as an important mediator for CTC immune evasion. Mechanistically, overexpression of CCL5 in CTCs is transcriptionally regulated by p38-MAX signaling, which recruites regulatory T cells (Tregs) to facilitate immune escape and metastatic seeding of CTCs. Collectively, our results reveal a previously unappreciated spatial heterogeneity and an immune-escape mechanism of CTC, which may aid in designing new anti-metastasis therapeutic strategies in HCC.

Structural and biological properties of polysaccharides from lotus root.

To investigate the structure, in vitro digestibility and activity of polysaccharides from lotus root, their main fractions named LRPs were isolated and purified by gel filtration chromatography. Structural analyses indicated that: LRPs were α­(1 → 6)­d­heteroglucans mainly composed of Glc-(1→, →6)-Glc-(1→, →6)-Gal-(1→, →4,6)-Gal-(1→ and →3,6)-Glc-(1→ at a molar ratio of 1.00: 4.33: 0.83: 0.13: 1.14; the total molar percentage of other monosaccharides in LRPs, including Man, Rha, GalA and Ara, was 8.10%; the molecular weights of LRPs was in the range of 1.33 kDa to 5.30 kDa. According to the change of molecular weight and the productions of reducing sugar and free monosaccharide, the simulated experiments of salivary, gastric and intestinal digestion confirmed that LRPs were almost undigestible. Moreover, LRPs showed the scavenging ability against DPPH and hydroxyl radicals, the growth inhibition ability against SGC7901 and HepG2 cancer cells in vitro, and the immunostimulating effect on the NO and TNF-α productions of macrophages in vitro. LRPs nearly remain their initial structure and activities in upper gastrointestinal tract and have health-improving potentials.

Comparison of stress response following microwave ablation and surgical resection of benign thyroid nodules.

PURPOSE: To compare the effects and safety of using microwave ablation (MWA) and surgical resection for the treatment of benign thyroid nodules (BTNs) under ultrasonic guidance and investigate the effects of this treatment on stress response. METHODS: Patients with BTNs were divided into the MWA and operation groups (72 cases each). Interleukin (IL)-6, IL-8, serum tumor necrosis factor (TNF-α), and hydrostatic visual analog scale (VAS) prior to the operation, at 6 h, 24 h, and 72 h post-operation were compared between the two groups. Operation times, hospitalization times, hospitalization expenses, and postoperative complications in the two groups were also compared. All patients underwent routine ultrasound and thyroid function testing at 3 and 6 months post-operation for assessment of nodule changes and thyroid hormone levels. RESULTS: Compared to the MWA group, the operation group had longer average operation times, longer hospital stays, a higher rate of neck pain after surgery, and a higher rate of fever (P < 0.05). Body temperature, as well as VAS, IL-6, IL-8, and TNF-α levels in the operation group were higher than those in the MWA group at 6 h, 24 h, and 72 h post-operation (P < 0.05). The levels of free thyroxine and free triiodothyronine in the operation group were lower than those in the MWA group (P < 0.05). CONCLUSION: MWA is a safe and effective treatment for patients with BTNs. The effects of MWA are more tolerable than those of surgical resection and the physiological function of the thyroid is preserved, which has high clinical value.

Intraoperative monitoring of pelvic autonomic nerves during laparoscopic low anterior resection of rectal cancer.

BACKGROUND: Some patients with low rectal cancer experience anorectal and urogenital dysfunctions after surgery, which can influence the long-term quality of life. In this study, we aimed to protect nerve function in such scenarios by performing intraoperative monitoring of pelvic autonomic nerves (IMPAN). PATIENTS AND METHODS: We retrospectively investigated a series of 87 patients undergoing laparoscopic low anterior resection of rectal cancer. Nerve-sparing was evaluated both visually and electrophysiologically. IMPAN was performed by stimulating the pelvic autonomic nerves under processed electromyography of the internal anal sphincter. Urination, defecation, sexual function, and the quality of life were evaluated using validated and standardized questionnaires preoperatively and at follow-up, 12 months after surgery. RESULTS: Among a total of 87 patients (53 male and 34 female patients), IMPAN with simultaneous electromyography of the internal anal sphincter was performed in 58 (66.7%) patients. Bilateral positive IMPAN results for both measurements, indicating successfully confirmed pelvic autonomic nerve preservation, were obtained in 45 (51.7%) patients. No significant difference was found in terms of urogenital and anorectal functions between preoperative and postoperative patients with bilateral positive IMPAN (P>0.05). Compared to preoperative patients with IMPAN (unilateral) or without IMPAN, these patients exhibited higher International Prostate Symptom Score, a lower International Index of Erectile Function-5, and a lower Female Sexual Function Index score at 12 months postoperatively (P<0.05). CONCLUSION: IMPAN is an appropriate method with which to laparoscopically protect nerve function.

Cytotoxic lanostane triterpenoids from the stems of Schisandra glaucescens.

Phytochemical investigation on the stems of Schisandra glaucescens resulted into the isolation of three new lanostane triterpenoids, 12-hydroxyschiglausin B (1), 12-hydroxykadsuphilactone B (2), and 20R-hydroxyschinalactone C (3). Structural elucidation of all the compounds was accomplished by spectral methods. The isolated compounds were tested in vitro for cytotoxic activities. As a result, triterpenoids 1 and 2 showed cytotoxic activities for all six tested tumor cell lines with IC50 values less than 15 µM.

HBx promotes the proliferative ability of HL­7702 cells via the COX­2/Wnt/ß­catenin pathway.

Hepatitis B virus X protein (HBx) has been termed a viral oncoprotein, and is involved in the initiation and progression of hepatocellular carcinoma (HCC). Cyclooxygenase­2 (COX­2) and ß­catenin have been attributed to the oncogenic activity of HBx in HBV­associated HCC. The present study aimed to determine whether there is crosstalk between COX­2 and the Wnt/ß­catenin signaling pathway during HL­7702­HBx cell proliferation, and to investigate the associated underlying molecular mechanism.  In the present study, cell proliferation assay, colony formation assay and flow cytometric analysis were used to detect the proliferative ability of cells. Reverse transcription­quantitative polymerase chain reaction and western blotting were performed to examine the mRNA and protein expression of COX­2, ß­catenin, cyclin­D1 and c­myc. The results demonstrated that HL­7702­HBx exhibited increased cell proliferation, higher colony formation efficiency and a shortened G1 period of the cell cycle. In addition, the mRNA and protein expression levels of COX­2 were increased, and this was associated with HL­7702­HBx cell growth. Furthermore, the expression of ß­catenin and its target genes, cyclin­D1 and c­myc proto­oncogene protein, was upregulated by HBx via COX­2. Finally, HBx promoted HL­7702 cell proliferation through the Wnt/ß­catenin signaling pathway. In conclusion, the primary finding of the present study was that HBx may promote HL­7702 cell proliferation via the COX­2/Wnt/ß­catenin pathway. Thus, it may be helpful to further investigate the molecular mechanism of HBV­associated hepatocellular carcinoma.

Hepatitis B Virus X protein elevates Parkin-mediated mitophagy through Lon Peptidase in starvation.

Hepatocellular Carcinoma (HCC) is the fifth most prevalent cancer worldwide. Specially, Hepatitis B viurs X protein (HBx) is a leading factor in the progression of Hepatitis B viurs-related HCC. Nutrient-deprived tumor microenvironment also contributes to tumor development. However, the role of HBx in nutrient-deprived HCC has received little investigation. Here, we show that HBx elevates PINK1-Parkin mediating mitophagy in starvation. HBx not only increases the PINK1/Parkin gene expression but also accelerates Parkin recruitment to partial mitochondria. Further analysis indicates that, HBx either promotes mitochondrial unfolded protein response, with remarkable mitochondrial LONP1 increases, or reduces LONP1 expression in cytosol inducing LONP1-Parkin pathway, both consequently enhancing mitophagy. Moreover, the enhanced mitophagy lowers mitochondrial apoptosis in starved hepatoma cells, and Bax is implied in the machinery. In addition, we define differential centrifuge, 3000 g or 12,000 g to pellet mitochondria, as an effective method to obtain distinct mitochondria. In collect, HBx regulates diverse aspects of LONP1 and Parkin, enhancing mitophagy in starvation. This study may shed new insights into the machinery development of hepatocellular carcinoma.

Association between FSP, CVHI, inflammatory cytokines and the incidence of primary stroke.

This case-control study was designed to establish a new risk-prediction model for primary stroke using Framingham stroke profile (FSP), cerebral vascular hemodynamic indexes (CVHI) and plasma inflammatory cytokines including hs-CRP, IL-6, TNF-α and Lp-PLA2. A total of 101 primary stroke patients admitted to Dongguan Houjie Hospital between August 2014 and June 2015 were assigned into the case group, and 156 age- and gender-matched healthy subjects from the Houjie Community were allocated into the control group. The prognostic values of FSP, CVHI and inflammatory cytokines including high sensitive C-reactive protein (hs-CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and lipoprotein-associated phospholipase A2 (Lp-PLA2) were assessed by multivariate logistic regression analysis. Seven risk-prediction models (FSP, CVHI, inflammatory cytokine, FSP+CVHI, FSP+inflammatory cytokine, CVHI+inflammatory cytokine, CVHI+FSP+inflammatory cytokine) were successfully established and the prognostic values were statistically compared by ROC curve and Z test. For FSP, the stroke risk was significantly elevated by 2.85 times when the FSP score was increased by 1 level (P=0.043), increased by 3.25 times for CVHI (P=0.036), 6.53 times for IL-6 (P=0.003), and 7.75 times for Lp-PLA2 (P=0.000). The sensitivity of FSP+CVHI+inflammatory cytokine and CVHI+inflammatory cytokine models was higher than 90%. For model specificity, the specificity of FSP+CVHI+inflammatory cytokine model alone exceeded 90%. FSP, CVHI, IL-6 and Lp-PLA2 are independent risk factors of stroke. Integrating IL-6 and Lp-PLA2 into the models can significantly enhance the risk prediction accuracy of primary stroke. Combined application of FSP+CVHI+inflammatory cytokine is of potential for risk prediction of primary stroke.

Monoterpenoid indole alkaloids from Alstonia mairei and their cytotoxicity.

Phytochemical investigation on the 70% ethanol extract of the leaves of Alstonia mairei resulted in the isolation of three new monoterpenoid indole alkaloids, alstomairines A-C (1-3), along with one known compound, alpneumine A (4). Structural elucidation of all the compounds was accomplished by spectral methods such as 1D and 2D NMR, IR, UV, and HRESIMS. The isolated compounds were tested in vitro for cytotoxic activities against four osteosarcoma cell lines. Consequently, alkaloids 2 and 3 exhibited cytotoxic activities for all tested tumor cell lines with IC50 values from 9.2 to 13.0 µM.

Hepatitis B virus X protein sensitizes HL-7702 cells to oxidative stress-induced apoptosis through modulation of the mitochondrial permeability transition pore.

Chronic hepatitis B virus (HBV) infection is a leading cause of liver cirrhosis and cancer. Among the pathogenic factors of HBV, HBV X protein (HBx) is attracting increased attention. Although it is documented that HBx is a multifunctional regulator that modulates cell inflammation and apoptosis, the exact mechanism remains controversial. In the present study, we explored the effect of HBx on oxidative stress-induced apoptosis in normal liver cell line, HL-7702. Our results showed that the existence of HBx affected mitochon-drial biogenesis by modulating the opening of the mitochondrial permeability transition pore (MPTP). Notably, this phenomenon was associated with a pronounced translocation of Bax from the cytosol to the mitochon-dria during the period of exposure to oxidative stress with a release of cytochrome c and activation of cleaved caspase-3 and PARP. Moreover, MPTP blockage with cyclosporin A prevented the translocation of Bax, and inhibited oxidative stress-induced apoptotic killing in the HBx-expressing HL-7702 cells. Our findings suggest that HBx exhibits pro-apoptotic effects upon normal liver cells following exposure to oxidative stress by modulating the MPTP gateway.

Amaryllidaceae alkaloids from the bulbs of Lycoris radiata with cytotoxic and anti-inflammatory activities.

Four new Amaryllidaceae alkaloids, (+)-1-hydroxy-ungeremine (1), (+)-6ß-acetyl-8-hydroxy-9-methoxy-crinamine (2), (+)-2-hydroxy-8-demethyl-homolycorine-α-N-oxide (3), (+)-N-methoxylcarbonyl-2-demethyl-isocorydione (4), together with two known compounds, (+)-6ß-acetyl-crinamine (5) and 8-demethyl-homolycorine-α-N-oxide (6) were isolated from the ethanol extract of the bulbs of Lycoris radiata. Structural elucidation of all the compounds were performed by spectral methods such as 1D and 2D ((1)H-(1)H COSY, HMQC, and HMBC) NMR spectroscopy, in addition to high resolution mass spectrometry. All the isolated alkaloids were in vitro evaluated for their cytotoxic activities against eight tumor cell lines (BEN-MEN-1, CCF-STTG1, CHG-5, SHG-44, U251, BGC-823, HepG2 and SK-OV-3) and anti-inflammatory activities against Cox-1 and Cox-2. As a result, alkaloids 1 and 4 exhibited significant cytotoxic activities against all tested tumor cell lines except against BEN-MEN-1. Additionally, alkaloids 1 and 4 possessed selective inhibition of Cox-2 comparable with the standard drug NS-398 (>90%).

Effectiveness and complications of anthracycline and taxane in the therapy of breast cancer: a meta-analysis.

OBJECTIVE: To compare the efficacy and safety between anthracycline & taxane and anthracycline in the treatment of breast cancer. METHODS: Computer-assisted literature search was performed with PubMed, MEDLINE, EMBASE and Cochrane Controlled Trials Register (CCTR) to identify pertinent literatures. Software RevMan 5.0 was used for statistical analysis. The measurement of interest outcomes included severe neurotoxicity, death without breast cancer recurrences, leukemia, venous thrombus and severe cardiotoxicity. RESULTS: A total of 10 randomized controlled trial studies (RCTs) containing 18,198 cases were selected in this meta-analysis. Of which, 9,902 cases were treated with anthracycline & taxane and 8,296 cases treated with anthracycline alone as control. Anthracycline & taxane showed lower risks of incident leukemia (RR = 0.40; 95% CI: 0.18, 0.90), venous thrombus (RR = 0.49; 95% CI: 0.29, 0.84) and severe cardiotoxicity (RR = 0.41, 95%CI: 0.26, 0.66), but higher risks of incident severe neurotoxicity (RR = 5.97; 95% CI: 1.72, 20.65) and non-recurrent death (RR = 1.79; 95% CI: 1.06, 3.04), compared to anthracycline alone. CONCLUSION: Clinically important differences exist for general safety in favour of the anthracycline & taxane rather than anthracycline alone. However, as a result of tumor recurrent rate, anthracycline might be superior to anthracycline & taxane. A longer duration of follow-up and a larger number of cases are required to better assess the efficacy and safety profile of the treatment of breast cancer.