Shenfu injection prevents sepsis-induced myocardial injury by inhibiting mitochondrial apoptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Shenfu injection (SFI) is a well-known Chinese herbal medicine widely used in the treatment of septic shock in China. AIMS: The aims of this study are to investigate the protective effects of SFI on sepsis-induced myocardial injury in mice and to identify the underlying mechanism of action. MATERIALS AND METHODS: Seventy-two male C57/B6J mice (5-6 weeks old) were randomly divided into five groups: control (NC), sham sepsis (sham), sepsis (Lipopolysaccharide- LPS), sepsis treated with a low dose SFI, and sepsis treated with a high dose SFI. Sepsis was induced in mice by intraperitoneal injection of LPS. Myocardial tissue samples were collected from different groups at 6 h, 12 h, and 24 h post-LPS injection. Myocardial injury was examined using hematoxylin-eosin (H&E) and TUNEL staining. Western-blot analysis was performed to determine the protein expression of B-cell lymphoma 2 (Bcl-2), BH3 interacting-domain death agonist (Bid), truncated-Bid (t-Bid) and caspase-9 in all the groups. Moreover, the structural changes in the mitochondria of cardiomyocytes were also observed by transmission electron microscopy. RESULTS: H&E staining revealed structural damage, local necrosis, interstitial edema, inflammatory cell infiltration and vacuolar changes in the myocardial tissue in the sepsis (LPS) group; almost intact myocardial tissue was observed in the high dose SFI group with improvements in interstitial edema and inflammatory cell infiltration. We observed that LPS-induced cardiomyocyte apoptosis was significantly improved with high dose SFI as compared with sepsis (LPS) group (P ˂ 0.05). LPS was found to decrease the protein expression of Bcl-2 and increase the level of Bid, t-Bid and caspase-9. Treatment with SFI significantly increased the Bcl-2 protein expression (P ˂ 0.05) and decreased the protein expression of Bid, t-Bid and caspase-9 as compared with LPS group (P ˂ 0.05). Markedly swollen myocardial mitochondria with partial vacuolation were observed in LPS treated mice while SFI treatment was found to significantly improve the LPS-induced morphological damage of the mitochondria. CONCLUSION: In conclusion, we demonstrate that SFI protects against sepsis-induced myocardial injury in mice through the suppression of myocardial apoptosis. It upregulates the protein expression of Bcl-2 and downregulates the protein expression of Bid, t-Bid and caspase-9, and alleviates sepsis-induced mitochondrial damage.

An automatic detection method for 40-Hz auditory steady state response and its application in prognosis of comatose patients.

OBJECTIVE: We proposed a 40-Hz auditory steady-state response (ASSR) automatic detection method, and studied the prognosis of comatose patients by combining the 40-Hz ASSR detection results of multiple paradigms of auditory stimulation. METHODS: The 40-Hz ASSR elicitation experiments were carried out on 32 comatose patients, with the detection results used as prognosis predictors. To achieve automatic detection, the detection was modeled as a binary hypothesis test for a sinusoidal waveform with unknown amplitude and phase, based on the generalized likelihood ratio test (GLRT). The patients were followed up for 6 months, and each patient's outcome was classified as either favorable outcome (severe disability, moderate disability or good recovery) or unfavorable outcome (vegetative state/unresponsive wakefulness syndrome or death) according to the Glasgow outcome scale (GOS). The performance of the prognosis predictors was assessed using the area under the receiver operating characteristic curve (AUC-ROC). RESULTS: The largest AUC in univariate analysis involving a single stimulation paradigm was 0.849, while the AUC obtained by combining multiple predictors was increased to 0.966. CONCLUSIONS: For comatose patients, the absence of 40-Hz ASSR in multiple stimulation paradigms may indicate an unfavorable prognosis. Furthermore, the combination of multiple auditory stimulation paradigms may increase the outcome prediction accuracy. SIGNIFICANCE: The combination of multi-paradigm 40-Hz ASSR automatic detection results may provide a feasible automatic outcome prediction method for comatose patients.

The correlation between the methylation of PTEN gene and the apoptosis of osteosarcoma cells mediated by SeHA nanoparticles.

The invasive spreading of residual osteosarcoma cells becomes a serious threat to human health, urgently needing new bone regenerative biomaterials for orthopedic therapy. Thus, in this work, selenite-substituted hydroxyapatite (SeHA) nanoparticles were prepared for both inhibiting the recurrence of the tumor and accelerating the regenerative repair of bone defect. Physicochemical characterization showed these synthetic nanoparticles were spherical poly-crystals with the shape of snowflakes. Such structure benefited them to inhibit the cellular viability of osteosarcoma cells by about (58.90 ± 14.37)% during 24 h co-culturing. The expression level of cell growth-related genes such as PTEN, MMP-9, Cyclin D1, Cyclin A2, Annexin A2 and CDC2 decreased. Bisulfite Sequence PCR of PTEN gene exhibited about (22.40 ± 5.39)%, (45.91 ± 6.36)% and (25.90 ± 5.36)% promoter methylation in control, HA and SeHA group. Animal experiment also proved the similar effects. Almost no recurrence were observed in SeHA group. Oppositely, the slowly recurrent growth of the remnant tumor appeared in purely surgical group. The overall survival and toxicity analysis showed that, in the usage dose of 0-0.1 g, the SeHA-0.01 exhibited higher inhibitory recurrence and metastasis potentials, lower renal toxicity and better anti-inflammation function. Immunohistochemistry stain showed the reduced expression of PTEN, MMP-9, Ki-67 and Annexin A2, but slightly increased expression of DNMT1 and BMP-2. Compared the methylation status of PTEN gene in each group, it was confirming that SeHA nanoparticles hardly possessed the de-methylation effect, but the pure HA strikingly increased the methylation level of such gene. It seemed the dopant selenite ions possessed de-methylation effect onto PTEN gene. Therefore, from the viewpoint of inhibiting metastatic potentials, the SeHA-0.01 might be a feasible biomaterial to inhibit the relapse of the tumor post-surgery.