Spinal microglial activation promotes perioperative social defeat stress-induced prolonged postoperative pain in a sex-dependent manner.

Prolonged postsurgical pain, which is associated with multiple risk factors in the perioperative stage, is a common medical and social problem worldwide. Suitable animal models should be established to elucidate the mechanisms underlying the perioperative prolonged postsurgical pain. In this study, standard and modified social defeat stress mice models, including chronic social defeat stress (CSDS), chronic nondiscriminatory social defeat stress (CNSDS) and vicarious social defeat stress (VSDS), were applied to explore the effect of perioperative social defeat stress on postsurgical pain in male and female mice. Our results showed that exposure to preoperative CSDS could induce prolonged postsurgical pain in defeated mice regardless of susceptibility or resilience differentiated by the social interaction test. Similar prolongation of incision-induced mechanical hypersensitivity was also observed in both sexes upon exposing to CNSDS or VSDS in the preoperative period. Moreover, we found that using the modified CNSDS or VSDS models at different recovery stages after surgery could still promote abnormal pain without sex differences. Further studies revealed the key role of spinal microglial activation in the stress-induced transition from acute to prolonged postoperative pain in male but not female mice. Together, these data indicate that perioperative social defeat stress is a vital risk factor for developing prolonged postoperative pain in both sexes, but the promotion of stress-induced prolonged postoperative pain by spinal microglial activation is sexually dimorphic in mice.

Triterpenoids and meroterpenoids from the edible Ganoderma resinaceum and their potential anti-inflammatory, antioxidant and anti-apoptosis activities.

Ganoderma resinaceum, as a traditional edible mushroom, has been widely reported to improve neurodegenerative diseases characterized by oxidative stress and inflammation. In this study, five new terpenoids, including four lanostane triterpenoids, named ganoresinoid A-D (1-4) and one meroterpenoid, named ganoresinoid E (5), along with 27 known compounds (6-32), were isolated from the fruiting bodies of edible mushroom G. resinaceum. These structures were identified by NMR, HRESIMS data analysis. All metabolites were evaluated for anti-inflammatory, antioxidative and anti-apoptosis activities. Among them, ganoresinoid A showed notably restrained nitric oxide (NO), IL-1ß, IL-6 and TNF-α levels in LPS-activated BV-2 microglial cells via suppressing TLR-4/ NF-κB and MAPK signaling pathway. Simultaneously, ganoresinoid A remarkably alleviated LPS-induced apoptosis by means of the decrease of mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). In addition, ganoresinoid A demonstrated antioxidant effects in H2O2-induced SH-SY5Y cells by activating the Akt/GSK-3ß/Nrf2 signaling pathway. Taken together, these results may provide a stronger theoretical basis for ganoresinoid A from G. resinaceum as nutrition intervention to alleviate neurodegenerative diseases.

[Construction and identification of gene vector expressing PTEN while simultaneously silencing Livin].

OBJECTIVE: To study the effect of simultaneously increasing PTEN gene expression and inhibiting Livin gene expression on the gastric carcinoma cell line (BGC823) and construct a recombinant vector expressing PTEN while simultaneously silencing Livin. METHODS: The siRNA expression unit against Livin gene (siLivin) was cleaved from pRNAT-U6.1-Livin vector and then inserted into pCL-neo-PTE to construct the recombinant vector pCL-neo-PTEN-siLivin. Then pCL-neo-PTEN-siLivinp, pCL-neo-PTEN, pRNAT-U6.1-Livin, pCL-neo and pRNAT-U6.1 were respectively transfected into the gastric carcinoma cell line (BGC823) with LipofectAMINE(TM) 2000. The mRNA and protein expression level of PTEN and Livin genes in each cell group was detected by fluorescent quantitative RT-PCR and Western blot. RESULTS: Recombinant vectors of pCL-neo-PTEN, pRNAT-U6.1-Livin and pCL-neo-PTEN-siLivin were constructed successfully. After transfection with pCL-neo-PTEN-siLivin, the mRNA and protein expression level of PTEN (0.897±0.112) rose in BGC823 cells while Livin gene became silenced. And the characterization of regulated cell bioactivity improved. There were significant differences between transfected and control groups (P<0.05). And the inhibiting effect on the proliferation and metastasis of BGC823 cell by increasing PTEN expression and silencing Livin simultaneously was better than that only by regulating PTEN genes or Livin genes alternatively. CONCLUSION: The recombinant vector of expressing PTEN and silencing Livin gene simultaneously is successfully constructed.

Characterization of cluster of differentiation 47 expression and its potential as a therapeutic target in esophageal squamous cell cancer.

The increased expression of cluster of differentiation (CD)47 has been identified in a number of different tumor types and is recognized as an adverse prognostic factor that indicates an increased risk of mortality in patients. The binding of CD47 to signal regulatory protein α (SIRPα) inhibits the macrophage phagocytosis of tumor cells by triggering an inhibitory 'do not eat me' signal. This is one of the mechanisms used by tumor cells to evade immune surveillance. In the present study, CD47 levels and macrophage infiltration were assessed in patients with esophageal squamous cell cancer (ESCC). CD47-overexpressing ESCC cell lines were selected and human M2 macrophage phagocytic activity was measured. The results revealed that CD47 is highly expressed and macrophages are markedly infiltrated in cancerous tissue compared with non-cancerous tissue. High CD47 expression was detected in ESCC cell lines and the results of a phagocytosis assay indicated that human M2 macrophages phagocytized tumor cells in a dose-dependent manner following the blocking of CD47-SIRPα signaling by anti-CD47 antibodies. The results of the present study therefore support the use of anti-CD47 immunotherapy to treat patients with ESCC.

Concomitant modulation of PTEN and Livin in gastric cancer treatment.

Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and Livin are important in the development of gastric cancer (GC). PTEN and Livin are involved in the regulation of tumor cell proliferation, migration and apoptosis. The modulation of PTEN or Livin has been investigated extensively in various cancer models. However, no studies have been performed to evaluate the combined effect of concurrently modulating these two genes on the development of GC. In the present study, the BGC823 human gastric carcinoma cell line was transfected with a dual gene modified vector (pCL-neo-PTEN-siLivin) in parallel with single gene modified vectors (pCL­neo­PTEN or pRNAT­U6.1­siLivin), and an empty control vector. Dual gene modulation (pCL­neo­PTEN­siLivin) had a more marked effect on the inhibition of cell proliferation, induction of apoptosis, and reduction of cell penetration in Matrigel, compared with either single gene alone or empty vector transfection. In a xenograft nude mouse model, the inoculation of pCL­neo­PTEN­siLivin­transfected BGC823 cells led to a markedly reduced tumor burden, compared with that in all other inoculation groups. In conclusion, the overexpression of PTEN concomitant with Livin gene silencing was confirmed as a feasible and effective in vitro and in vivo gene modulation method, which may represent a potential therapeutic strategy for the treatment of GC.

[Study on apoptosis effect induced by isothiocyanates in broccoli on HepG-2 cells and its mechanism].

OBJECTIVE: To investigate the apoptosis effect of isothiocyanates (ITCS) on human liver cancer cells HepG-2, and its mechanism. METHOD: HepG-2 cells were treated with different concentrations of ITCS. MTT assay was used to evaluate the influence of ITCS on cell proliferation. Flow cytometry was used to test ROS levels, intracellular mitochondrial transmembrane potential (deltapsim) , and hypodiploid apoptosis peak in HepG-2 cells. RESULT: ITCS obviously inhibited proliferation of HepG-2 cells. When treated with 15, 30, 60, 120, 240 microg x mL(-1) of ITCS for 24 h, ROS levels were (23.1+/-1. 8)%, (53.3+/-3.3)%, (57.9+/-2.0)%, (79.9+/-3.4)%, (93.4+/-2. 6)% respectively; and deltapsim were (94.8+/-5.5)%, (91.8+/-5.4)%, (66.0+/-5.6)%, (65. 5+/-6.6)% and (44.3+/-2.7)% respectively; when treated with 60, 120, 240 microg x mL(-1) of ITCS for 48 h, cell apoptotic rates were (16.6+/-2.8)%, (21.9+/-4.4) % and (70.2+/-5.3) % respectively. CONCLUSION: ITCS generates ROS in gastric cancer HepG-2 cells, which causes mitochondrial membrane permeabilization and deltapsim decrease, therefore, leads to apoptosis of HepG-2 cells.

The underlying cellular mechanism in the effect of tetramethylpyrazine on the anion secretion of colonic mucosa.

The present study investigated the underlying cellular mechanism in the effect of ligustrazine (tetramethylpyrazine, TMP) on the anion secretion of colonic mucosa in rats using a short-circuit current (I(sc)) technique in conjunction with "tool drugs." (i) After a pretreatment of the tissues by bathing the bilateral surface with Cl(-)-free Krebs-Henseleit (K-H) solution for over an hour, a basolateral application of 1 mmol/l TMP produced an increase in I(sc), and the total charges transported for 30 min were about 8.7 +/- 1.4 mC/cm(2); an apical pretreatment of DPC and a basolateral addition of acetazolamide decreased the TMP-induced I(sc) by about 60% (P < 0.01) and 45% (P < 0.05), respectively; a basolateral application of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), the inhibitor of Na(+)-HCO(3)(-) cotransporter (NBC), did not alter the TMP-induced I(sc). (ii) After the bilateral surface of mucosa was bathed with HCO(3)(-)-free K-H solution for over an hour, a basolateral application of 1 mmol/l TMP produced an increase in I(sc), and the total charges transported in 30 min were about 8.3 +/- 1.9 mC/cm(2); an apical pretreatment of DPC (1 mmol/l), the inhibitor of Cl(-) channels, decreased the TMP-induced Isc by about 84% (P < 0.01). The basolateral presence of bumetanide (0.1 mmol/l), the inhibitor of Na(+)-K(+)-Cl(-) cotransporter (NKCC), significantly reduced the TMP-evoked I(sc) by about 86% (P < 0.01). In conclusion, (i) ligustrazine could promote colonic mucosa secretion Cl(-) via apical Cl(-) channels and basolateral NKCC; (ii) ligustrazine could promote colonic mucosa secretion HCO(3)(-) via apical Cl(-) channels and the basolateral diffusion of CO(2).