[Correlation of serum and seminal plasma HCY levels with semen parameters in men and its effect on recurrent spontaneous abortion].

OBJECTIVE: To investigate the correlation of serum and seminal plasma homocysteine (Hcy) levels with semen parameters in men and its effect on recurrent spontaneous abortion (RSA) in their spouses. METHODS: The study included 103 males subjects undergoing preconception examination in the reproduction center from March 2022 to June 2023. According to whether their spouses had a history of RSA or not, we divided their subjects into an RSA (n = 43) and a non-RSA group (NRSA, n = 60), obtained their serum and seminal plasma Hcy levels and semen parameters, and analyzed their correlation. RESULTS: The serum Hcy level was significantly correlated with the sperm DNA fragmentation index (DFI) (r = 0.316, P = 0.005), but not with the seminal plasma Hcy level (r = －0.041, P = 0.723) and other semen parameters of the subjects (P > 0.05). There was no significant correlation between seminal plasma Hcy and semen parameters (P > 0.05). The median serum Hcy was significantly higher in the RSA than in the NRSA group (18.39 ï¼»13.02, 42.84ï¼½ vs 14.65 ï¼»12.00, 18.20ï¼½ µmol/L), with statistically significant difference in the overall distribution of serum Hcy between the two groups (Ｚ=－2.20, P = 0.028), so was the median sperm DFI in the former than in the latter group (25.00% ï¼»12.50%, 37.25%ï¼½ vs 13.00% ï¼»11.00%, 18.50%ï¼½), with statistically significant difference in the overall sperm DFI distribution between the two groups (Ｚ=－2.74, P = 0.006). CONCLUSION: The serum Hcy level was positively correlated with sperm DFI, and both serum Hcy and sperm DFI were significantly elevated in men with spousal RSA, which is expected to be used as a clinical screening indicator for males with spousal RSA.

[Oxalis inhibits prostate tumor in the mouse model of castration-resistant prostate cancer: Effect and mechanism].

OBJECTIVE: To investigate the inhibitory effect of oxalis on prostate tumor in the mouse model of castration-resistant prostate cancer (CRPC) and its action mechanism. METHODS: We established a CRPC model in 40 male C57/BL mice aged 6－8 weeks, divided them randomly into four groups of an equal number, and treated them intragastrically with normal saline (control), low-dose oxalis (5 mg/kg/d), medium-dose oxalis (10 mg/kg/d), and high-dose oxalis (15 mg/kg/d), respectively. After 28 days of treatment, we measured the tumor volume and body weight of the mice in different groups, calculated the tumor-inhibition rate, examined the histomorphological changes of the prostate tumors by HE staining, and detected the expressions of the nuclear factor-κB (NF-κB) signaling pathway and its downstream proteins in the tumor tissue by immunofluorescence assay. RESULTS: In comparison with the controls, the mice in the low-, medium- and high-dose oxalis groups showed a gradual decrease in tumor cell concentration and cell degeneration, and a gradually increased number of necrotic tumor cells. The volume and mean weight of prostate tumors were significantly reduced (P < 0.05), the expressions of NF-κB p65 and Ki67 proteins remarkably down-regulated (P < 0.05), and that of the Bax protein markedly up-regulated (P < 0.05) in the oxalis groups compared with the controls. CONCLUSION: Oxalis can inhibit the growth of prostate tumor in CRPC mice possibly by down-regulating the NF-κB signaling pathway and the expressions of p65 and Ki67 and up-regulating the expression of Bax, and thereby promoting the degeneration and necrosis of tumor cells.

[Mechanism of regulation of CNP rat model by oxalis decoction via cGAS-STING signaling pathway].

OBJECTIVE: To investigate the therapeutic mechanism of oxalis decoction on CNP rats by regulating cGAS-STING signaling pathway. METHODS: Thirty specific pathogen-free SD male rats were randomly divided into normal control group (NC), model control group (MC), and oxalis decoction group (OD),with 10 rats in each group.The left and right anterior abdominal lobes of each group were surgically exposed.The normal control group was injected by the same volume of normal saline.After the model was successfully established,the OD group was given ï¼»9.37g/(kg·d)ï¼½ by gavage once a day, and the NC and MC groups were given ï¼»0.01/(mｌ/g)ï¼½ normal saline by gavage. From the 7th day of administration, the body weight of the rats in each group was recorded every 7 days for dynamic comparison. After 50 days of administration, the prostate index of the rats in each group was calculated, the morphological and pathological changes of the prostate tissue were observed by HE staining,and the expression levels of tumor necrosis factorα(TNF-α), interleukin-1ß(IL-1ß)and IL-6 in serum were detected by ELISA. RT-qPCR was used to detect the mRNA expression of cGAS, STING, TRAF6 and HSP70 in prostate tissue of rats in each group. RESULTS: Versus the NC group and OD group, the prostate organ index in MC group was significantly higher than that in other groups (P<0.01). Versus the NC group, the HE staining results of the MC group showed that the prostate gland structure was disordered, and the interstitial and acinar epithelium were extensively edema, accompanied by a large number of lymphocyte infiltration, cell swelling, loose cytoplasm, and a small number of foam cells. Versus the MC group, HE staining showed that the edema of interstitial and acinar epithelial cells in the rat prostate tissue was reduced after the OD group intervention, and the inflammatory cell infiltration in the interstitium was significantly reduced.Versus to NC group, the expression levels of TNF-α,IL-1ßandIL-6 in MC group were significantly increased（P<0.01 ).Versus to MC group,the expression levels of TNF-α, IL-1ß and IL-6 in OD group were decreased (P<0.05). Versus the NC group, the mRNA expression of cGAS, STING and TRAF6 in the MC group was significantly up-regulated,and HSP70mRNA was significantly down-regulated(P<0.01).Versus the MC group,the OD group had significantly decreased mRNA expression of cGAS, STING and TRAF6 and significantly increased mrna expression of HSP70(P<0.05). CONCLUSIONS: CNP has autoimmune disorders that cause inflammatory responses.The key target for CNP treatment is to regulate innate immunity.The treatment with oxalis decoction can significantly improve the prostate organ index and pathological changes in CNP rats, which may be related to the down-regulation of cGAS-STING innate immune signaling pathway and the inhibition of inflammatory mediators secretion.

[Action mechanisms of Xiaoluowan (II) in the treatment of epididymitis: A network pharmacology-based study].

OBJECTIVE: To explore the potential action mechanisms of Xiaoluowan (II) (XLW-II) in the treatment of epididymitis through a network pharmacology approach. METHODS: We searched various databases for relevant targets associated with epididymitis and XLW-II and obtained the common targets of epididymitis and XLW-II on the Venny platform. We acquired the protein-protein interactions (PPI) using the STRING data and had them visualized with the Cytoscape software. After topological analysis, we retrieved the key targets, followed by gene ontology (GO) and KEGG pathway enrichment analyses using the DAVID database. RESULTS: A total of 2 38 drug targets, 2 150 disease targets and 85 common targets were identified. The core targets for the treatment of epididymitis with XLW-II identified by PPI network analysis included TNF, IL6, IL1B, MMP9, AKT1, PTGS2 and TP53. GO function analysis revealed the involvement of the common targets in such biological processes as response to hypoxia, regulation of apoptotic processes, inflammatory response, and positive regulation of the MAPK cascade. KEGG pathway analysis suggested that the signaling pathways such as the cancer pathway, PI3K-Akt pathway, protein glycosylation pathway in cancer, Ras pathway and chemokine pathway might be related to the action mechanisms of XLW-II in the treatment of epididymitis. CONCLUSION: The potential targets and signaling pathways of Xiaoluowan (II) in the treatment of epididymitis were identified on the basis of network pharmacology, which has provided a novel insight into its action mechanisms and offered a new direction for further relevant studies.

[Exploring the mechanism of levocarnitine in the treatment of epididymitis based on network pharmacology and molecular docking technology].

OBJECTIVE: To explore the potential mechanism of action of levocarnitine in the treatment of epididymitis based on network pharmacology and experimental research. METHODS: The target proteins related to epididymitis and levocarnitine were retrieved through multiple databases, and the common targets were obtained using Venny software. The protein-protein interactions were obtained using the STRING database. Cytoscape software was used for visualization, and key targets were selected after topological analysis. GO and KEGG pathway enrichment analysis was performed using the DAVID database. Molecular docking was performed using Autodock Vina. RESULTS: A total of 130 drug targets and 2 151 disease targets were obtained, with 47 common targets. Protein-protein interaction network analysis identified core targets of levocarnitine in the treatment of epididymitis, including AKT1, HSP90AA1, ALB, CASP3, GSK3B, and GSR. KEGG pathway analysis suggested that metabolic pathways, lipid metabolism and atherosclerosis, cancer pathways, fluid shear stress and atherosclerosis, measles, chemical carcinogens-reactive oxygen species, purine metabolism, PI3K-Akt, and other signaling pathways may be associated with the mechanism of levocarnitine in the treatment of epididymitis. CONCLUSION: This study revealed through network pharmacology that levocarnitine may act on multiple signaling pathways by targeting AKT1, HSP90AA1, ALB, CASP3, GSK3B, GSR, etc., thereby potentially exerting therapeutic effects on epididymitis.

[Research on the mechanism of Miao ethnicity medicine formula of Oxalis corniculata against chronic non-bacterial prostatitis].

Objective： To explore the mechanism of Miao ethnicity medicine formula of Oxalis corniculata against chronic non-bacterial prostatitis. Methods: The rat model of chronic abacterial prostatitis was induced by stimulation with 2% sterile carrageenan solution. After modeling, the rats were randomly divided into two groups, Model control group (Model group) and oxalis group. Another normal control group was set up. The rats in the Model group and the normal control group were given 0.01ml/g normal saline by gavage, and the rats in the oxalis alis group were given 1ml/100g (1 g/kg) of Oxalis corniculata L warm water decoction by gavage once a day for 28 days. Twenty-four hours after the last administration, 10ml blood was collected from the abdominal aorta of the rats, and prostate tissue samples were collected from the rats. hematoxylin-eosin (HE) staining was used to observe the morphological structure of the prostate in normal and prostatitis rats. ELISA was used to detect the levels of serum and prostate cytokines. The protein expressions of 4-HNE , ALDH2 and FGF2 were detected by Western blot. Results: Compared with the blank group, the model group showed obvious hyperplasia of fibrous tissue in the interstitium of the prostate tissue, disordered glandular structure, papillary hyperplasia of epithelial cells in the acini, infiltration of a small amount of lymphocytes, monocytes and other inflammatory cells in the acini, and increased pathological scores. The protein expressions of 4-HNE , ALDH2 , MCP-1 and FGF2 in prostate tissue were significantly increased. Compared with the model group, the prostate tissue of the oxalis group was slightly damaged, with a small amount of fibrous hyperplasia and inflammatory cell infiltration. The protein expressions of 4-HNE , ALDH2 , MCP-1 and FGF2 were decreased in prostate tissue. Conclusion： Oxalis corniculata L can effectively repair the pathological morphology of prostate tissue in rats with CNP, and its mechanism may be related to activating 4-HNE protein and reducing oxidative stress injury of prostate tissue in rats.

[Biological mechanisms of Oxalis corniculata regulating human prostate cancer PC-3 cells: An investigation based on the NF-κB pathway].

OBJECTIVE: To investigate the biological mechanisms underlying the effect of the Chinese herbal medicine Oxalis corniculata on human prostate cancer PC-3 cells. METHODS: Through in vitro experiment, we treated human prostate cancer PC-3 cells with different concentrations of Oxalis corniculata, assessed the viability of the cells by MTT assay, examined their apoptosis by flow cytometry, evaluated their migration and invasiveness by Transwell assay, and determined the expressions of the proteins p65, p-p65, IκBα and p-IκBα in the NF-κB pathway using protein imprinting technology. RESULTS: Compared with the blank control, Oxalis corniculata significantly inhibited the proliferation and induced the apoptosis of the PC-3 cells (P< 0.05), suppressed their migration and invasiveness in a dose-dependent manner (P< 0.05), and upregulated the expression of IκBα and downregulated those of p-p65 and p-IκBα in the NF-κB pathway (P< 0.05). CONCLUSION: Oxalis corniculata can inhibit the proliferation, migration and invasiveness and induce the apoptosis of human prostate cancer PC cells, which may be attributed to its abilities of inhibiting the expressions of p-p65 and p-IκBα and regulating the activity of the NF-κB pathway.

[Mechanism of Xiaoluanwan(II) in treating mouse epididymitis through the regulation of NLRP3 inflammasome].

OBJECTIVE: This study aimed to investigate the mechanism of Xiaoluanwan(II) in treating lipopolysaccharide(LPS)-induced epididymitis and its impact on the NLRP3 inflammasome. METHODS: The murine epididymitis model was established through local injection of LPS. The study included a control group (n=5), a model group (n=5), a model group treated with Xiaoluanwan(II) (Ⅱ) (n=5), and a saline group treated with Xiaoluanwan(II) (n=5). After 14 consecutive days of oral administration of Xiaoluanwan(II) or physiological saline, pathological changes in the epididymal tissues, expression levels of NLRP3 inflammasome and Caspase-1, as well as associated protein levels were examined. RESULTS: Compared to the model group, Xiaoluanwan(II) significantly alleviated inflammatory cell infiltration and lesions, as evidenced by a reduction in the protein expression levels of NLRP3, Caspase-1, Cleaved-Caspase-1, IL-1ß, IL-18, GSDMD, and p-p38 MAPK (P<0.05 or P<0.01), thereby mitigating the inflammatory response. CONCLUSION: Xiaoluanwan(II) alleviates epididymal inflammation and ameliorates mouse epididymal epithelial injury by modulating the NLRP3-mediated cell pyroptosis pathway.

Inhibition of hepatocyte nuclear factor 1ß contributes to cisplatin nephrotoxicity via regulation of nf-κb pathway.

Cisplatin nephrotoxicity has been considered as serious side effect caused by cisplatin-based chemotherapy. Recent evidence indicates that renal tubular cell apoptosis and inflammation contribute to the progression of cisplatin-induced acute kidney injury (AKI). Hepatocyte nuclear factor 1ß (HNF1ß) has been reported to regulate the development of kidney cystogenesis, diabetic nephrotoxicity, etc However, the regulatory mechanism of HNF1ß in cisplatin nephrotoxicity is largely unknown. In the present study, we examined the effects of HNF1ß deficiency on the development of cisplatin-induced AKI in vitro and in vivo. HNF1ß down-regulation exacerbated cisplatin-induced RPTC apoptosis by indirectly inducing NF-κB p65 phosphorylation and nuclear translocation. HNF1ß knockdown C57BL/6 mice were constructed by injecting intravenously with HNF1ß-interfering shRNA and PEI. The HNF1ß scramble and knockdown mice were treated with 30 mg/kg cisplatin for 3 days to induce acute kidney injury. Cisplatin treatment caused increased caspase 3 cleavage and p65 phosphorylation, elevated serum urea nitrogen and creatinine, and obvious histological damage of kidney such as fractured tubules in control mice, which were enhanced in HNF1ß knockdown mice. These results suggest that HNF1ß may ameliorate cisplatin nephrotoxicity in vitro and in vivo, probably through regulating NF-κB signalling pathway.

[CCL2 promotes angiogenesis of primary rat cardiac microvascular endothelial cells].

The aim of the present study was to investigate the role of chemokine CCL2 in angiogenesis of primary adult rat cardiac microvascular endothelial cells (CMEC). The rat CMECs were isolated and identified through morphology examination and immunostaining with CD31 and factor VIII antibodies. The angiogenesis of CMEC on Matrigel was evaluated at different time points. The expression and secretion of CCL2 during the process of angiogenesis was detected by real-time RT-PCR and ELISA, respectively. The results showed that, the primary rat CMEC was isolated successfully, and the angiogenesis of CMEC was significantly induced after Matrigel treatment for 4 h. The expression of CCL2 and CCR2 were increased during angiogenesis, and the secretion of CCL2 was detected after 2 h of angiogenesis and reached the peak concentration of 1 588.1 pg/mL after 4 h. Either CCL2 blocking antibody or CCR2 antagonist significantly reduced the angiogenesis of CMEC. These results suggest that CCL2 is secreted during the process of angiogenesis of CMEC, and CCL2/CCR2 signaling pathway may play an important role in promoting angiogenesis.

RNA interference may suppress stress granule formation by preventing argonaute 2 recruitment.

RNA-induced silencing complex (RISC) is formed during RNA interference (RNAi), whereas stress granules (SG) are assembled in response to cellular stress. Here, we demonstrate an interesting connection between RISC and SG that may involve argonaute 2 (Ago2), a core component of RISC. We analyzed SG induction by arsenite, the commonly used SG inducer. SG formation was suppressed in heat shock transcription factor 1 (Hsf1) or hypoxia-inducible factor-1α (Hif1α) shRNA-transfected cells but not in Hsf1 or Hif1α-knockout cells, suggesting that RNAi per se (rather than gene deficiency) may account for the suppressive effect on SG. In support, the suppressive effect of RNAi on SG formation was reversed by the RISC-loading inhibitor aurintricarboxylic acid. In non-RNAi cells, arsenite induced the accumulation of Ago2 in SGs as shown by its colocalization and coimmunoprecipitation with SG proteins, but Ago2 was not recruited to SG in the cells with RNAi. Consistently, arsenite induced the dissociation of Ago2 from RISC proteins in non-RNAi cells but not in RNAi cells. CRISPR-Cas9-medicated ablation of Ago2 attenuated SG formation during arsenite treatment, suggesting a critical role of Ago2 in SG assembly. Together, these results indicate that RISC and SG may compete for some key components, such as Ago2. In response to cellular stress, Ago2 is recruited for SG assembly; however, during RNAi, Ago2 is held in RISC, becoming unavailable for SG formation.

MicroRNA-375 Is Induced in Cisplatin Nephrotoxicity to Repress Hepatocyte Nuclear Factor 1-ß.

Nephrotoxicity is a major adverse effect of cisplatin-mediated chemotherapy in cancer patients. The pathogenesis of cisplatin-induced nephrotoxicity remains largely unclear, making it difficult to design effective renoprotective approaches. Here, we have examined the role of microRNAs (miRNAs) in cisplatin-induced nephrotoxicity. We show that cisplatin nephrotoxicity was not affected by overall depletion of both beneficial and detrimental miRNAs from kidney proximal tubular cells in mice in which the miRNA-generating enzyme Dicer had been conditionally knocked out. To identify miRNAs involved in cisplatin nephrotoxicity, we used microarray analysis to profile miRNA expression and identified 47 up-regulated microRNAs and 20 down-regulated microRNAs in kidney cortical tissues. One up-regulated miRNA was miR-375, whose expression was also induced in cisplatin-treated renal tubular cells. Interestingly, inhibition of miR-375 decreased cisplatin-induced apoptosis, suggesting that miR-375 is a cell-damaging or pro-apoptotic agent. Blockade of P53 or NF-κB attenuated cisplatin-induced miR-375 expression, supporting a role of P53 and NF-κB in miR-375 induction. We also identified hepatocyte nuclear factor 1 homeobox B (HNF-1ß) as a key downstream target of miR-375. Of note, we further demonstrated that HNF-1ß protected renal cells against cisplatin-induced apoptosis. Together, these results suggest that upon cisplatin exposure, P53 and NF-κB collaboratively induce miR-375 expression, which, in turn, represses HNF-1ß activity, resulting in renal tubular cell apoptosis and nephrotoxicity.

Hsf4 counteracts Hsf1 transcription activities and increases lens epithelial cell survival in vitro.

The interplay between Hsf4 and Hsf1 plays an important role in the regulation of lens homeostasis. However, the mechanism of the intermolecular association involved is still unclear. In this paper, we find that reconstitution of Hsf4b into Hsf4-/- lens epithelial (mLEC/Hsf4-/-) cells can simultaneously downregulate Hsp70 expression and upregulate the expression of small heat shock proteins Hsp25 and αB-crystallin at both RNA and protein levels. ChIP assay results indicate Hsf4b, which binds to the promoters of Hsp90α, Hsp70.3, Hsp25 and αB-crystallin but not Hsp70.1, can inhibit Hsf1 binding to Hsp70.3 promoter and the heat shock mediated Hsp70 promoter activity by reducing Hsf1 protein expression. Hsf4b N-terminal hydrophobic region can interact with Hsf1 N-terminal hydrophobic region. Their interaction impairs Hsf1's intramolecular interaction between the N- and C-terminal hydrophobic regions, leading to Hsf1's cytosolic retention and protein degradation. Both lysosome inhibitors (chloroquine, pepstatin A plus E64d) and proteasome inhibitor MG132 can inhibit Hsf4-mediated Hsf1 protein degradation, but MG132 can induce Hsf1 activation as well. Upregulation of Hsf4b can significantly inhibit cisplatin and staurosporine induced lens epithelial cell apoptosis through direct upregulation of Hsp25 and αB-crystallin expression. Taken together, our results imply that upregulation of Hsf4b modulates the expression pattern of heat shock proteins in lens tissue by either directly binding to their promoters or promoting Hsf1 protein degradation. Moreover, upregulation of Hsf4b protects lens cell survival by upregulating anti-apoptotic pathways. These studies reveal a novel regulatory mechanism between Hsf1 and Hsf4b in modulating lens epithelial cell homeostasis.

Heat shock factor 1 induces crystallin-αB to protect against cisplatin nephrotoxicity.

Cisplatin, a wildly used chemotherapy drug, induces nephrotoxicity that is characterized by renal tubular cell apoptosis. In response to toxicity, tubular cells can activate cytoprotective mechanisms, such as the heat shock response. However, the role and regulation of the heat shock response in cisplatin-induced nephrotoxicity remain largely unclear. In the present study, we demonstrated the induction of heat shock factor (Hsf)1 and the small heat shock protein crystallin-αB (CryAB) during cisplatin nephrotoxicity in mice. Consistently, cisplatin induced Hsf1 and CryAB in a cultured renal proximal tubular cells (RPTCs). RPTCs underwent apoptosis during cisplatin treatment, which was increased when Hsf1 was knocked down. Transfection or restoration of Hsf1 into Hsf1 knockdown cells suppressed cisplatin-induced apoptosis, further supporting a cytoprotective role of Hsf1 and its associated heat shock response. Moreover, Hsf1 knockdown increased Bax translocation to mitochondria and cytochrome c release into the cytosol. In RPTCs, Hsf1 knockdown led to a specific downregulation of CryAB. Transfection of CryAB into Hsf1 knockdown cells diminished their sensitivity to cisplatin-induced apoptosis, suggesting that CryAB may be a key mediator of the cytoprotective effect of Hsf1. Taken together, these results demonstrate a heat shock response in cisplatin nephrotoxicity that is mediated by Hsf1 and CryAB to protect tubular cells against apoptosis.

Two-component signal transduction system SaeRS is involved in competence and penicillin susceptibility in Staphylococcus epidermidis.

Staphylococcus epidermidis, which is a causative pathogen of nosocomial infection, expresses its virulent traits such as biofilm and autolysis regulated by two-component signal transduction system SaeRS. In this study, the S. epidermidis SaeRS was identified to negatively regulate the expression of genes involved in competence (comF, murF), cytolysis (lrgA), and autolysis (lytS) by DNA microarray or real-time RT-PCR analysis. In addition, saeRS mutant showed increased competence and higher susceptibility to antibiotics such as penicillin and oxacillin than the wild-type strain. The study will be helpful for understanding the characterization of the SaeRS in S. epidermidis.

Regulation of Hsf4b nuclear translocation and transcription activity by phosphorylation at threonine 472.

Hsf4b, a key regulator of postnatal lens development, is subjected to posttranslational modifications including phosphorylation. However, the phosphorylation sites in Hsf4b and their biological effects on the transcription activity of Hsf4b are poorly understood. Here we examined 17 potential phosphorylation residues in Hsf4b with alanine-scanning assays and found that a T472A mutation diminished Hsf4b-mediated expression of Hsp25 and alphaB-crystallin. In contrast, the phosphomimetic mutation of T472D enhanced their expression. Further investigation demonstrated that Hsf4b could interact with nuclear-transporter importin beta-1 and Hsc70 via amino acids 246-320 and 320-493, respectively. T472A mutation reduced Hsf4bs interaction with importin beta-1, while enhancing its interaction with Hsc7O, resulting in Hsf4b cytosolic re-localization, protein instability and transcription activity attenuation. At the upstream, MEK6 was found to interact with Hsf4b and enhance Hsf4b's nuclear translocation and transcription activity, probably by phosphorylation at sites such as T472. Taken together, our results suggest that phosphotylation of Hsf4b at T472 by protein kinases such as MEI(6 regulates Hsf4b interaction with the importin V I -Hsc7O complex, resulting in blockade of its nuclear translocation and transcriptional activity of Hsf4b.

Glucose regulates heat shock factor 1 transcription activity via mTOR pathway in HCC cell lines.

HSF1-mediated heat shock response is activated in most tumors and plays important roles in regulating tumor homeostasis. However, the signals underlying HSF1 activation is still not completely understood. In this paper, we find that glucose, the dominant tumor energy supplement, participates in regulating HSF1's activation in HCC cell lines. The immunoblotting results indicate that the phosphorylation of HSF1/S326, a hallmark of HSF1 activation, varies between the HCC cell lines (e.g., SMMC7721, HapG2, plc/prf5, and Chang-liver). Glucose, but not 2D-glucose, can induce the phosphorylation of HSF1 at S326 and upregulate the expression of HSF1's downstream alpha B-crystallin and Hsp70 as well as the none-heat shock proteins CSK2 and RBM23 in two tested hepatocellular carcinoma cell lines (prl/prf5 and SMMC7721). Rapamycin, an inhibitor of mTOR, can suppress the glucose-induced phosphorylation of HSF1/S326 and the expression of alpha B-crystallin. Knockdown of HSF1 with shRNA enhances the glucose-depletion-mediated inhibition of plc/prf5 cell proliferation. Our data reveal that HSF1 can be activated by glucose-mTOR pathway, providing an alternative pathway for targeting HSF1 in tumor therapy.

[Regulatory effect of two component signal transduction system saeRS on the survival of Staphylococcus epidermidis].

OBJECTIVE: To investigate the survival ability of Staphylococcus epidermidis (S. epidermidis) wildtype (WT), saeRS mutant (SAE) and saeRS complementary (SAEC) strains under different concentrations of glucose. METHODS: We measured the survival ability, biofilm forming ability, medium acidity and antimicrobial susceptibility of S. epidermidis in medium containing different concentrations of glucose. RESULTS: Compared with WT, the survival ability, biofilm forming ability and resistance to antibiotics (such as penicillin, oxacillin, gentamicin, ciprofloxacin and amikacin) of saeRS mutant increased significantly in response to glucose. SAE and SAEC showed the strongest survival ability and biofilm forming ability when grown in medium containing 14 mmol/L glucose and 28 mmol/L, respectively. WT showed no significant different survival and biofilm forming abilities when cultured with various concentrations of glucose. The medium acidity of saeRS mutant (pH=8.07) was lower than the WT (pH=7.0) in the presence of 14 mmol/L glucose. CONCLUSION: SaeRS may influence the survival ability of S. epidermidis by affecting glucose utilization.

Proteomic profiling of N-linked glycoproteins identifies ConA-binding procathepsin D as a novel serum biomarker for hepatocellular carcinoma.

The aim of this study was to identify novel biomarkers for the diagnosis of, and potential therapeutic targets for, hepatocellular carcinoma (HCC). Multilectin affinity chromatography was used to enrich N-linked glycoproteins from nontumorous liver and HCC tissues followed by 2DE and protein identification by MS. Twenty-eight differentially expressed proteins were identified. Western blotting validated consistently lower concentrations of human liver carboxylesterase 1 and haptoglobin, and higher concentration of procathepsin D (pCD) in HCC tissues. Knockdown of cathepsin D (CD) expression mediated by siRNA significantly inhibited the in vitro invasion of two HCC cell lines, SNU449 and SNU473, which normally secrete high-levels of CD. Prefractionation using individual lectins demonstrated an elevation in ConA-binding glycoforms of proCD and CD in HCC tissues. In the serum of HCC patients, "ConA-binding proCD" (ConA-pCD) is significantly increased in concentration and this increase is comprised of several distinct upregulated acidic isoforms (pI 4.5-5.5). Receiver operating characteristic analysis showed that the sensitivity and specificity of serum ConA-pCD for HCC diagnosis were 85% and 80%, respectively. This is the first report that serum ConA-pCD is increased significantly in HCC and is potentially useful as a serological biomarker for diagnosis of HCC.

Two-component signal transduction system SaeRS positively regulates Staphylococcus epidermidis glucose metabolism.

Staphylococcus epidermidis, which is a causative pathogen of nosocomial infection, expresses its virulent traits such as biofilm and autolysis regulated by two-component signal transduction system SaeRS. In this study, we performed a proteomic analysis of differences in expression between the S. epidermidis 1457 wild-type and saeRS mutant to identify candidates regulated by saeRS using two-dimensional gel electrophoresis (2-DE) combined with matrix-assisted laser desorption/lonization mass spectrometry (MALDI-TOF-MS). Of 55 identified proteins that significantly differed in expression between the two strains, 15 were upregulated and 40 were downregulated. The downregulated proteins included enzymes related to glycolysis and TCA cycle, suggesting that glucose is not properly utilized in S. epidermidis when saeRS was deleted. The study will be helpful for treatment of S. epidermidis infection from the viewpoint of metabolic modulation dependent on two-component signal transduction system SaeRS.