Yunming Li

Notch1 and Notch2 have opposite prognostic effects on patients with colorectal cancer

BACKGROUND Aberrantly activated Notch signaling has been shown to play a key role in carcinogenesis and progression of various human malignancies. In this study, we investigated the expression of Notch1 and Notch2 in colorectal cancer to determine whether they could serve as prognostic predictors. PATIENTS AND METHODS The protein expression of Notch1 and Notch2 was examined by immunohistochemistry in 1003 clinical colorectal cancer specimens. Notch1 and Notch2 protein levels were investigated by immunohistochemistry. Statistical analysis was carried out to assess their prognostic value. RESULTS Significantly negative correlation between Notch1 and Notch2 was found in colorectal cancer (P < 0.001). Notch1 and Notch2 were proved to be inversely correlated with tumor differentiation, depth of invasion, lymph node metastases, distant metastasis, TNM (tumor-node-metastasis) stage and survival of patients, suggesting opposite function of the two receptors. Notch1 and Notch2 were proved to be adverse independent prognostic predictors (P < 0.001). Moreover, a synergistic effect of positive Notch1 and negative Notch2 coexpression on predicting poor overall survival was proved. CONCLUSIONS Notch1 and Notch2 may be independent adverse prognostic predictors for patients with colorectal cancer. These results would contribute to identify more efficient prognostic predictors and therapeutic targets.

Hypoxic preconditioning attenuates global cerebral ischemic injury following asphyxial cardiac arrest through regulation of delta opioid receptor system.

This study was designed to investigate whether delta opioid receptor (DOR) is involved in the neuroprotective effect induced by hypoxic preconditioning (HPC) in the asphyxial cardiac arrest (CA) rat model. Twenty-four hours after the end of 7-day HPC, the rats were subjected to 8-min asphyxiation and resuscitated with a standardized method. In the asphyxial CA rat model, HPC improved the neurological deficit score (NDS), inhibited neuronal apoptosis, and increased the number of viable hippocampal CA1 neurons at 24 h, 72 h, or 7 days after restoration of spontaneous circulation (ROSC); however, the above-mentioned neuroprotection of HPC was attenuated by naltrindole (a selective DOR antagonist). The expression of hypoxia-inducible factor-1α (HIF-1α) and DOR, and the content of leucine enkephalin (L-ENK) in the brain were also investigated after the end of 7-day HPC. HPC upregulated the neuronal expression of HIF-1α and DOR, and synchronously elevated the content of L-ENK in the rat brain. HIF-1α siRNA was used to further elucidate the relationship between HIF-1α and DOR in the HPC-treated brain. Knockdown of HIF-1α by siRNA markedly abrogated the HPC induced upregulation of HIF-1α and DOR. The present study demonstrates that the expression of DOR in the rat brain is upregulated by HIF-1α following exposure to 7-day HPC, at the same time, HPC also increases the production of endogenous DOR ligand L-ENK in the brain. DOR activation after HPC results in prolonged neuroprotection against subsequent global cerebral ischemic injury, suggesting a new mechanism of HPC-induced neuroprotection on global cerebral ischemia following CA and resuscitation.

Astrocytic activation in thoracic spinal cord contributes to persistent pain in rat model of chronic pancreatitis

One of the most important symptoms in chronic pancreatitis (CP) is constant and recurrent abdominal pain. However, there is still no ideal explanation and treatment on it. Previous studies indicated that pain in CP shared many characteristics of neuropathic pain. As an important mechanism underlying neuropathic pain, astrocytic activation is probably involved in pain of CP. Based on the trinitrobenzene sulfonic acid (TNBS)-induce rat CP model, we performed pancreatic histology to assess the severity of CP with semiquantitative scores and tested the nociceptive behaviors following induction of CP. Glial fibrillary acidic protein (GFAP) expressions in the thoracic spinal cord were observed by immunohistochemistry and real-time reverse transcription polymerase chain reaction (RT-PCR). Meanwhile, we injected intrathecally astrocytic specific inhibitor l-alpha-aminoadipate (LAA) and observed its effect on nociception induced by CP. Compared to the naive and sham group, TNBS produced long lasting pancreatitis, and persistent mechanical hypersensitivity in the abdomen that was evident 1 week after TNBS infusion and persisted up to 5 weeks. Compared with naive or sham operated rats, GFAP staining was significantly increased 5 weeks after CP induction. Real-time RT-PCR indicated that GFAP expression was significantly increased in TNBS treated rats compared to the sham group. TNBS-induced astrocytic activation was significantly attenuated by LAA, compared with the saline control. Treatment with LAA significantly, even though not completely, attenuated the allodynia. Our results provide for the first time that astrocytes may play a critical role in pain of CP. Some actions could be taken to prevent astrocytic activation to treat pain in CP patients.

Matrix Metalloproteinase-9 Is Associated with Relapse and Prognosis of Patients with Colorectal Cancer

BackgroundMatrix metalloproteinase-9 is a member of the MMP family, which is overexpressed in some solid tumors and is thought to enhance tumor invasion and metastasis ability. The present study aims to examine MMP-9 expression in human colorectal cancer and to determine its association with clinicopathological characteristics and prognosis.MethodsColorectal cancer and adjacent normal tissues from 192 patients were investigated by immunohistochemical assay. Staining evaluation results were analyzed statistically in relation to various clinicopathological characters, disease-free survival, and overall survival.ResultsHigh level of MMP-9 expression was detected in colorectal cancer, significantly more than in normal colorectal epithelial cells. In colorectal cancer, MMP-9 was significantly positively correlated with depth of invasion, lymph node metastasis, and distant metastasis. However, no correlations between MMP-9 expression and patient age, sex, tumor location or differentiation status were detected. Disease-free and overall survival were significantly poorer for patients with positive MMP-9 staining than for those with MMP-9-negative tumors.ConclusionsOur findings emphasize the important role of MMP-9 in the invasion and metastasis process in human colorectal cancer. It could also serve as a novel prognostic marker that is independent of, and additive to, the tumor–node–metastasis (TNM) staging system.

Notch2 Expression Is Decreased in Colorectal Cancer and Related to Tumor Differentiation Status

BackgroundThe significance of Notch2 expression in colorectal cancer (CRC) has not been clearly investigated. We investigated the expression of Notch2 and its relationship with differentiation status and tumor stage by studying clinical CRC specimens with matched adjacent normal tissues and normal control colorectal specimens.MethodsImmunohistochemistry, real-time polymerase chain reaction, and Western blot analysis were performed to assess the expression of Notch2 in clinical CRC specimens. Also, Notch2 levels in an induced differentiation model of CRC cell lines were investigated.ResultsIt was found that Notch2 expression was decreased in cancer tissues compared with adjacent normal tissue and normal control tissues. Also, a tendency for decreased expression was observed when going from well to poorly differentiated carcinomas, as well as going from tumor, node, metastasis system stage I to stage IV. With the differentiation of colon cancer cells, the expression of Notch2 increased. To support this observation, colon cancer cell lines HT29 and SW620 were induced to differentiate in culture, and expression of Notch2 was investigated. A clear increase expression of Notch2 was observed.ConclusionsNotch2 expression correlated closely with CRC and may play a role in tumor inhibition in colon carcinogenesis.

Increased MicroRNA-630 Expression in Gastric Cancer Is Associated with Poor Overall Survival

MicroRNAs are noncoding RNAs that regulate multiple cellular processes during cancer progression. Among various microRNAs, MiR-630 has recently been identified to be implicated in many critical processes in human malignancies. We aimed to investigate the significance and prognostic value of miR-630 in human gastric cancer. Gastric cancer and adjacent normal specimens from 236 patients from who had not received neoadjuvant chemotherapy were collected. The expression of miR-630 was investigated by quantitative real-time PCR assay and its association with overall survival of patients was analyzed by statistical analysis. MiR-630 expression level was significantly elevated in gastric cancer in comparison to adjacent normal specimens. It is also proved that miR-630 expression was to be associated with gastric cancer invasion, lymph node metastasis, distant metastasis and TNM stage. In addition, survival analysis proved that elevated miR-630 expression was associated with poor overall survival of patients. Multivariate survival analysis also proved that miR-630 was an independent prognostic marker after adjusted for known prognostic factors. The present study proved the over-expression of miR-630 and its association with tumor progression in human gastric cancer. It also provided the first evidence that miR-630 expression was an independent prognostic factor for patients with gastric cancer, which might be a potential valuable biomarker for gastric cancer.

Effects of intracerebroventricular application of the delta opioid receptor agonist [d-Ala2, d-Leu5] enkephalin on neurological recovery following asphyxial cardiac arrest in rats

The delta opioid receptor (DOR) agonist [D-Ala2, D-Leu5] enkephalin (DADLE) has been implicated as a novel neuroprotective agent in the CNS. The current study was designed to evaluate the effects of intracerebroventricular (ICV) application of DADLE on neurological outcomes following asphyxial cardiac arrest (CA) in rats. Male Sprague-Dawley rats were randomly assigned to four groups: Sham group, CA group, DADLE group (DADLE+CA), and Naltrindole group (Naltrindole and DADLE+CA). All drugs were administered into the left cerebroventricle 30 min before CA. CA was induced by 8-min asphyxiation and the animals were resuscitated with a standardized method. DOR protein expression in the hippocampus was significantly increased in the CA group at 1 h after restoration of spontaneous circulation (ROSC) compared with the Sham group. As time progressed, expression of DOR proteins decreased gradually in the CA group. Treatment with DADLE alone or co-administration with Naltrindole reversed the down-regulation of DOR proteins in the hippocampus induced by CA at 24 h after ROSC. Compared with the CA group, the DADLE group had persistently better neurological functional recovery, as assessed by neurological deficit score (NDS) and Morris water maze trials. The number of surviving hippocampal CA1 neurons in the DADLE group was significantly higher than those in the CA group. However, administration of Naltrindole abolished most of the neuroprotective effects of DADLE. We conclude that ICV administration of DADLE 30 min before asphyxial CA has significant protective effects in attenuating hippocampal CA1 neuronal damage and neurological impairments, and that DADLE executes its effects mainly through DOR.

Notch1 Expression in Colorectal Carcinoma Determines Tumor Differentiation Status

IntroductionThe significance of Notch1 expression in colorectal cancer has not been clearly described. We investigated the expression of Notch1 and its relationship with differentiation status and tumor (Union Internationale Contre le Cancer, UICC) stage using a series of 237 colorectal cancer samples with matched adjacent normal tissues and a series of 46 normal colorectal specimens.Materials and MethodsImmunohistochemistry, real-time polymerase chain reaction, and Western blot analysis were performed to assess the expression of Notch1.ResultsIt was found that Notch1 was overexpressed in cancer tissues as compared with adjacent normal tissue and normal control tissues. Also, a tendency for increased expression was observed when going from well to poorly differentiated carcinomas, as well as going from UICC stage I to stage IV. With the differentiation of colon cancer cells, the expression of Notch1 decreased. To support this observation, colon cancer cell lines HT29 and SW620 were induced to differentiate in culture, and expression of Notch1 was investigated. A clear reduction of Notch1 expression was observed.ConclusionThese results suggest that Notch1 expression correlated closely with colorectal cancer and may play an oncogenic role during colonic carcinogenesis.

The reciprocal connections of endomorphin 1- and endomorphin 2-containing neurons between the hypothalamus and nucleus tractus solitarii in the rat.

In the CNS, endomorphin 1- and endomorphin 2-immunoreactive neuronal cell bodies have been principally found both in the hypothalamus and nucleus tractus solitarii. Functionally, the hypothalamus and nucleus tractus solitarii are closely related in many aspects, especially in visceral functions. On the other hand, there are also many endomorphin-immunoreactive fibers and terminals in the two regions. In the present study, to investigate whether endomorphin 1-immunoreactive and endomorphin 2-immunoreactive neurons in the hypothalamus and nucleus tractus solitarii project reciprocally between these two regions, fluorescent retrograde labeling combined with immunofluorescence histochemical staining for endomorphin 1 and endomorphin 2 was used. After injection of Fluoro-Gold into the nucleus tractus solitarii of rats, endomorphin 1/Fluoro-Gold or endomorphin 2/Fluoro-Gold double-labeled neuronal cell bodies were predominantly observed in the arcuate nucleus of the hypothalamus, a few of which were also observed in the posterior hypothalamic area and periventricular hypothalamic nucleus. After injection of Fluoro-Gold into the medial zone of hypothalamic tuberal region and the lateral hypothalamic area, respectively, endomorphin 1/Fluoro-Gold or endomorphin 2/Fluoro-Gold double-labeled neuronal cell bodies were found chiefly in the medial, commissural, lateral and gelatinous parts of the nucleus tractus solitarii. These results provide morphological evidence that there exist reciprocal endomorphinergic connections between the hypothalamus and nucleus tractus solitarii.

The role of 5-HT receptor subtypes in the ventrolateral orbital cortex of 5-HT-induced antinociception in the rat

The present study examined the involvement of 5-HT in the ventrolateral orbital cortex (VLO) on descending antinociception and determined which subtypes of 5-HT receptors mediated this effect. This study focused on the effects of 5-HT microinjection in the VLO of lightly anesthetized male rats on the radiant heat-evoked tail flick (TF) reflex, as well as the influence of 5-HT(1A), 5-HT(2), 5-HT(3), and 5-HT(4) receptor subtype antagonists on the effect of 5-HT. Results showed that 5-HT microinjection (2, 5, 10 microg, in 0.5 microl) into the VLO depressed the TF reflex in a dose-dependent manner. Pretreatment with 5-HT receptor antagonists (1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine hydrobromide (NAN-190), cyproheptadine hydrochloride (CPT) and 1-methyl-N-(8-methyl-8-azabicyclo[3.2.3]-oct-3-yl)-1H-indazole-3-carboxamide maleate salt (LY-278,584)), specific for 5-HT(1A), 5-HT(2) and 5-HT(3) receptors, respectively, partially reversed the 5-HT-evoked inhibition. In contrast, the 5-HT(4) receptor antagonist, 1-[2-[(methylsulfonyl)-amino]ethyl]-4-piperidinyl]methyl1-methyl-1H-indole-3-carboxylate (GR 113808), had no effect on the inhibition of 5-HT. Microinjections of NAN-190, CPT and LY-278,584 alone into the VLO had no effect on the TF reflex. These results suggest that 5-HT(1A), 5-HT(2) and 5-HT(3), but not 5-HT(4) receptors, are involved in mediating 5-HT-induced antinociception in the VLO. According to different properties and distribution patterns of the 5-HT receptor subtypes on neurons, the possible mechanism of 5-HT activation of the VLO-periaqueductal gray (PAG) descending antinociceptive pathway is discussed.