Potential of semen coicis in enhancing the anti-tumor effects of PD-1 inhibitor on A549 cell lines by blocking the PI3K-AKT-mTOR pathway.

BACKGROUND: The objective of this research was to investigate how the combination of semen coicis extract and PD-1 inhibitors can potentially work together to enhance the anti-tumor effects, with a focus on understanding the underlying mechanism. METHODS: We obtained the active components and specific targets of semen coicis in the treatment of NSCLC from various databases, namely TCMSP, GeneCard, and OMIM. By utilizing the STRING database and Cytoscape software, we established a protein interaction network (PPI) for the active ingredient of semen coicis and the target genes related to NSCLC. To explore the potential pathways involved, we conducted gene ontology (GO) and biological pathway (KEGG) enrichment analyses, which were further supported by molecular docking technology. Additionally, we conducted cyto-inhibition experiments to verify the inhibitory effects of semen coicis alone or in combination with a PD-1 inhibitor on A549 cells, along with examining the associated pathways. Furthermore, we investigated the synergistic mechanism of these two drugs through cytokine release experiments and the PD-L1 expression study on A549 cells. RESULTS: Semen coicis contains two main active components, Omaine and (S)-4-Nonanolide. Its primary targets include PIK3R1, PIK3CD, PIK3CA, AKT2, and mTOR. Molecular docking experiments confirmed that these ingredients and targets form stable bonds. In vitro experiments showed that semen coicis demonstrates inhibitory effects against A549 cells, and this effect was further enhanced when combined with PD-1 inhibitors. PCR and WB analysis confirmed that the inhibition of the PI3K-AKT-mTOR pathway may contribute to this effect. Additionally, semen coicis was observed to decrease the levels of IFN-γ, IL-6, and TNF-α, promoting the recovery of the human anti-tumor immune response. And semen coicis could inhibit the induced expression of PD­L1 of A549 cells stimulated by IFN­Î³ as well. CONCLUSION: Semen coicis not only has the ability to kill tumor cells directly but also alleviates the immunosuppression found in the tumor microenvironment. Additionally, it collaboratively enhances the effectiveness of PD-1 inhibitors against tumors by blocking the activation of PI3K-AKT-mTOR.

Regulation of the Paneth cell niche by exogenous L-arginine couples the intestinal stem cell function.

Although previous studies show that exogenous nutrients regulate the stem cell function, little is known about the effects of L-arginine on intestinal stem cells (ISCs). In this study, we utilize mice, small intestinal (SI) organoids, and ISC-Paneth cell co-cultured models to clarify the role of L-arginine in ISC function. We find that exogenous L-arginine is essential for ISCs proliferation and intestinal epithelial renewal. Our data show that Paneth cells, a critical component of the ISCs niche, augment the ISCs function in response to L-arginine. Moreover, enhanced the expression of Wnt3a in Paneth cells, which is a ligand of the Wnt/ß-catenin signaling pathway, mediates the effects of L-arginine on ISCs function. Pre-treatment with L-arginine enhances the ISCs pool and protects the gut in response to injury provoked by murine tumor necrosis factor α (TNF-α) and 5-Fluorouracil (5-FU). Our findings establish that the regulation of Wnt3a in the Paneth cell niche by exogenous L-arginine couples ISCs function and favours a model in which the ISCs niche couples the nutrient levels to ISCs function.

PSD95 gene specific siRNAs attenuate neuropathic pain through modulating neuron sensibility and postsynaptic CaMKIIα phosphorylation.

OBJECTIVE: To observe the effects of PSD95 gene specific siRNAs on neuropathic pain relief, neuron viability, and postsynaptic calcium/calmodulin-dependent protein kinase IIα (CaMKIIα) phosphorylation in vitro and in vivo. METHODS: Gene-specific siRNAs of rat PSD95 were synthesized chemically for transfection. Adult male Sprague-Dawley (SD) rats were randomly divided into 3 groups: naïve group (n=6), sham group (n=6), and sciatic nerve chronic constriction injury (CCI) group (n=24). The CCI group was further divided into 4 groups (n=6 in each group), which were pretreated with normal saline, transfection vehicle, negative control siRNAs, and PSD95 gene specific siRNAs respectively. All the subgroups received corresponding agents intrathecally for 3 days, started one day before the CCI of sciatic nerve. Both mechanical allodynia and thermal hyperalgesia were measured on post-operative day 3 and 7. PSD95 gene silenced NG108-15 cells were further stimulated by glutamate, with the cell viability and the expression/phosphorylation of CaMKIIα measured by MTT cell proliferation assay and Western blot, respectively. RESULTS: The siRNAs decreased PSD95 mRNA level significantly both in vivo and in vitro. Neuropathic pain rats pretreated with PSD95 gene specific siRNAs exhibited significant elevation in the mechanical withdrawal threshold and paw withdrawal thermal latency, without affecting the baseline nociception. PSD95 gene silencing enhanced neuronal tolerance against the glutamate excitotoxicity, meanwhile the phosphorylation of CaMKIIα Thr286 was attenuated. CONCLUSION: Pre-emptive administration of PSD95 gene specific siRNAs may attenuate the central sensitization CaMKIIα-related signaling cascades, leading to the relief of neuropathic pain.

[Expressions of NOS isoforms in the cavernous tissues of diabetic rat models].

OBJECTIVE: To investigate the mechanism of diabetic erectile dysfunction (ED) and find new methods for its treatment by detecting the changes in nitric oxide synthase (NOS) isoforms and erectile function of diabetic rats and observing the effects of insulin and alpha-lipoic acid (LA) on it. METHODS: Fifty male Sprague-Dawley rats were divided into Groups A (normal control, n=10), B (non-intervention diabetes mellitus, n=13), C (insulin intervention diabetes mellitus, n=12), and D (insulin + LA intervention, n=15). And the diabetic models were made by intraperitoneal injection of streptozocin (STZ). Eight weeks later, the erectile function of the rats was assessed following apomorphine injection and the contents of NOS isoforms in the erectile tissues measured by immunohistochemical staining. RESULTS: All the rats of Group A showed a normal erectile function (100%). In comparison, those in Groups B, C and D exhibited a significantly decreased rate, 28.6% in Group B, 62.5% in Group C and 80.9% in Group D. The numbers of positive nNOS fibers and eNOS in the penile tissues per visual field were 86.7 and 9.6 in Group A, but only 36.5 and 3.3 in Group B, 52.7 and 5.7 in Group C, and 71.4 and 7.4 in Group D (P < 0.05). However, the expression of iNOS was significantly lower in Group A (6.9) than in Groups B (43.6), C (36.2) and D (19.3) (P < 0.05). Compared with Groups B and C, the erectile function and the expressions of nNOS and eNOS were markedly increased, while the expression of iNOS significantly decreased in Group D (P < 0.05). CONCLUSION: Diabetes mellitus severely affects penile erectile function and the expressions of NOS isoforms in the cavernous tissues, for which hyperglycemia is mainly responsible. LA is proved obviously efficacious for diabetic ED, which might be related to its antioxidant effect.

[Expression of nerve growth factor in cavernous tissue and its effects on the treatment of rats with diabetic erectile dysfunction].

OBJECTIVE: To investigate the content of nerve growth factor (NGF) in penis of rats with diabetic erectile dysfunction (DED) and apply rhNGF to treat the DED rat model to study the pathogenesis mechanism of DED and treatment effects by NGF for DED rat model. METHODS: Fifty adult male SD rats were randomly selected to make up diabetic models. After 8 weeks, the mRNA and protein levels of NGF in rat penis were detected. Then the rest of rats were divided into 5 groups: normal control group, non-treated diabetes group, NGF treatment group, insulin treatment group, NGF plus insulin treatment group. After 8 weeks of treatment, intracavernous pressure (ICP) was measured. The content of neuronal nitric oxide synthase (nNOS) in the erectile tissue was also value- ated by immunohistochemistry staining. RESULTS: Compared with those of normal control group, the mRNA content of NGF and protein in the penis of non-treated diabetic rats is increased significantly. Compared with non-treated diabetic group, ICP was much higher in the treated groups which received NGF or/and insulin therapy, and the changes in nNOS staining of those groups were the same. CONCLUSION: The injury of pelvic splanchnic nerves in advanced stage of diabetes may lead to DED, which may be relevant to the abnormal of NGF level or the NGF receptors. The increased extent of NGF in penis of rat with DED suggests that erectile nerves be severely damaged by diabete and the increase of producing NGF couldn't compensate the needs for reproduction of the nerve fibers. The ability of combination of NGF with its receptor may also be damaged. It is helpful to use exogenous NGF to lessen the partly neuropathy and improve the erectile dysfunction of diabetic rats. The abnormal of NGF may play an important role in the pathogenesis of diabetic ED and its treatment.

[The results of surgery on HIV carriers with urinary system disease].

OBJECTIVE: To approach characteristics of performing operation on HIV carriers with urinary system diseases. METHODS: To summarize author's experiences of surgery on 41 HIV carriers suffering urinary system diseases abroad from April 1996 to May 2004. RESULTS: The 41 HIV carriers received HAART and were performed with corresponding operations, followed up from 4 to 30 months post-operatively. The 31 carriers have recovered well up to date, while 4 carriers died of AIDS. Among them, 2 patients with penis cancer who received a partial peotomy and a patient with renal tuberculosis receiving left nephrectomy were died of AIDS within 4-8 months after operations whose CD4+ T lymphocyte number was below 0.2 x 10(9)/L. CONCLUSION: Prior to operation, HIV carriers should receive HAART ordinarily to control copy of the virus. The CD4+ T lymphocyte number is important for selecting a proper time for operation and deciding the further after surgery. We also take note to CD4+ T lymphocyte number to monitor progress of the AIDS. For those HIV carriers, endourologic surgery and laparoscopy should be taken so far as possible. Meanwhile, medical stuffs must pay more attention to preventing occupational infection during surgery.