MiR-199 inhibits EMT and invasion of hepatoma cells through inhibition of Snail expression.

OBJECTIVE: MiR-199 expression is associated with liver cancer. Bioinformatics analysis revealed that miR-199 has a complementary binding site to the 3'-UTR region of Snail mRNA. This study investigated whether miR-199 plays a role in regulating Snail expression and affecting epithelial-mesenchymal transition (EMT) and invasion of hepatoma cells. PATIENTS AND METHODS: The Dual-Luciferase reporter gene assay validated the targeted regulation between miR-199 and Snail. QRT-PCR was used to detect and compare the expression of miR-199 and Snail mRNA in human normal liver HL7702 cells, low metastatic MHCC97L cells, and high metastatic MHCC97H cells. MHCC97H cells were cultured in vitro and divided into two groups: miR-NC group and the miR-199 mimic group followed by the analysis of the expression of Snail, E-cadherin, and N-cadherin, as well as cell invasion ability by transwell assay. RESULTS: There was a targeted regulatory relationship between miR-199 and Snail mRNA. Compared with HL7702 cells, miR-199 expression was significantly decreased, and Snail expression was significantly increased in MHCC97L and MHCC97H cells, with more changes being observed in high metastatic MHCC97H cells. The transfection of miR-199 mimic significantly downregulated the expression of Snail and N-cadherin in MHCC97H cells, increased E-cadherin expression, inhibited the cell's EMT process, and invasion. CONCLUSIONS: The decrease of miR-199 expression plays a role in upregulating the expression of Snail and promoting EMT and invasion of hepatocarcinoma cells. The increase of the expression of miR-199 can inhibit the expression of Snail and inhibit the EMT process and invasion ability of hepatoma cells.

[Clinical characteristics and treatment of myoepithelial carcinoma of head and neck].

Objective:To investigate the clinical features, diagnosis, treatment and prognosis of myoepithelial carcinoma of the head and neck. Method:The clinical data of 59 patients with head and neck myoepithelial carcinoma admitted from January 2012 to October 2018 were analyzed retrospectively. The data was analyzed with SPSS 21.0 statistical software. Result:The follow-up period was 6 to 131 months. The mean follow-up time was 36 months. One patient（1.7%） was lost to follow-up, 17 patients（28.8%） had postoperative local recurrence, 8 patients（13.6%） had distant metastasis, and 5 patients（8.5%） had cervical lymph node metastasis; 14 patients（23.7%） died. The 1-, 3-, and 5-year cumulative survival rates were 92%, 73%, and 62%, respectively. Survival rate curves of different treatment methods were significantly different by Gehan method（P<0.05）. Compared between the two groups, there was significant difference between surgery alone and surgery plus radiotherapy than radiotherapy alone and chemotherapy alone（P<0.05）. There was no significant difference between the other two treatment methods. Multivariate logistic regression analysis showed that tumor location, clinical stage and survival status and local recurrence rate were significantly correlated（P<0.05）, but gender, age and survival status and local recurrence rate were not significantly correlated（P>0.05）. Conclusion:The incidence of myoepithelial carcinoma is low, and the clinical manifestations and imaging studies lack specificity. The tumor is prone to local recurrence, invasive, and has a high incidence of distant metastasis. It is a highly malignant tumor. Surgical treatment is preferred and the requirements for first surgery are high, and major salivary glands and advanced tumors（stage Ⅲ-Ⅳ） are risk factors for survival and local recurrence. Early diagnosis and early treatment can significantly improve the survival rate of patients, reduce the local recurrence rate of tumors, and improve the prognosis of patients.

Perceptions of public attitudes towards persons with mental illness in Beijing, China: results from a representative survey.

OBJECTIVES: Many studies have examined the general public's attitudes towards people with mental illness, but such studies are scarce in China. This study examined the perceptions of the Beijing population regarding their society's prevalent attitudes towards people with mental illness. METHODS: A total of 5000 individuals aged 18 or above living in Beijing were selected using a multistage, stratified, cluster and random sampling method. This was followed by a face-to-face interview which used a standardized questionnaire asking about societal attitudes towards individuals with mental illness. RESULTS: 4602 out of 5000 eligible individuals met the inclusion criteria and participated in the interview. 4596 questionnaires were deemed valid and included in the analyses. A large proportion of respondents believed that most individuals within their society held negative attitudes and had a strong desire to distance themselves from people with mental illness. Respondents aged 60 or older, who lived farther to downtown Beijing, or with higher education tended to believe that most individuals have relatively positive and tolerant attitudes towards people with mental illness. CONCLUSIONS: Many people in Beijing perceive that most members of their society have negative beliefs towards people with mental illness. Further efforts are needed to determine if these perceptions are accurate and to reduce the stigma that is reinforced by these perceptions.

A complication of xanthogranulomatous cholecystitis with Mirizzi syndrome.

A patient had right upper quadrant pain with sclera was transferred from emergency room to the hospital, she was proposed to have acute cholecystitis, gallstones, obstructive jaundice, and a four-year history of gallbladder stones. The NMR results showed that the gallbladder was significantly enlarged and the gallbladder wall was thickening irregularly. The liver morphology was not abnormal except with extensive intrahepatic bile duct dilatation. The MRCP results demonstrated that the intrahepatic bile ducts were significant expanded. The ERCP results showed that duodenal stenosis and extra-hepatic bile duct stenosis. We placed a plastic stent of 8.5Fr and 12 cm in length in the hepatic duct, and after biliary plastic stent placement, jaundice was rapidly reduced and liver function was improved significantly. A surgery was performed and the final pathologic diagnosis is a complication of Xanthogranulomatous cholecystitis with Mirizzi syndrome. After the surgery of cholecystectomy and a bile duct repair were performed, the patient was recovered well. Conclusively, if a patient was diagnosed as biliary stricture, a biliary metal stent should not be placed until pathological diagnosis of malignancy.

Effect of orphanin FQ on interleukin-1beta mRNA transcripts in the rat CNS.

To elucidate the mechanism of orphanin FQ on neuroimmune modulation, the relationship between orphanin FQ and interleukin-1beta in the rat CNS in vivo and in vitro was investigated. In our experiments, it was found that orphanin FQ and interleukin-1beta mRNA transcripts showed a similar distribution in cerebral cortex, hippocampus and hypothalamus. By using the in situ hybridization technique, down-regulation of interleukin-1beta mRNA transcripts by central administration of orphanin FQ was further identified in the traumatic animal model. Similar inhibitory effects were also observed on the number of microglia in the CNS. The effects produced by orphanin FQ were abolished by combination with its receptor (OP(4))-specific antagonist [Phe(1)Psi(CH(2)-NH)Gly(2)]nociceptin-(1-13)-NH(2), which suggested that the function of orphanin FQ might be attributable to the OP(4) pathway. However, the effect on the number of astrocytes in the CNS remained unchanged, despite evidence that OP(4) is expressed on astrocytes as well as on neurons and microglia. When analyzed by reverse transcription-polymerase chain reaction, interleukin-1beta gene expression was observed to be enhanced and inhibited in primary neuron and microglial cell cultures exposed to orphanin FQ respectively. Interleukin-1beta gene expression in astrocyte cultures was not affected by treatment with orphanin FQ. Our findings suggest that the neuroimmune function of orphanin FQ might be dependent on interleukin-1beta derived from microglia, and the interaction between microglia and neurons.

Effects of electroacupuncture on blood-brain barrier after cerebral ischemia-reperfusion in rat.

The effect of electroacupuncture (EA) on the blood-brain barrier (BBB) after cerebral ischemia-reperfusion (I-R) was investigated in rats. The dye Evans Blue (EB) was used as a tracer for assessing the disruption of BBB. Fluorescence quantification of EB was performed to explore the temporal pattern of permeability of BBB after the cerebral I-R with a fluorescence spectrophotometer (HITACHI 650-60). Furthermore, the morphology of BBB opening was detected under confocal laser scanning microscopy system. It was found that the BBB opening after cerebral I-R was biphasic. A rare scattered extravasation of EB was detected 2 hours after cerebral I-R. The EB extravasation reached its first peak at 6 h then decreased at 24 h and increased again at the time-point of 48h after cerebral I-R. EA can attenuate the disruption of BBB after cerebral I-R. EA could not only limit the area of extravasation of EB, but also reduce the concentration of extravasation of EB in the rat brain after cerebral I-R. The results indicated that one of the mechanisms of curative effect of EA on the cerebral ischemia might be due to its function of protecting the integrity of BBB.

Alteration of orphanin FQ immunoreactivity and ppOFQ mRNA by combination of melatonin with electroacupuncture.

The aim of the present study was to observe the alternation of central orphanin FQ (OFQ, also known as nociceptin) system while electroacupuncture (EA) combined with melatonin (MEL). The experiments were carried out to investigate the changes of OFQ-like immunoreactivity and prepro-orphanin FQ (ppOFQ) mRNA in some certain nuclei of the rat brain. Using immunohistochemical technique we found that the level of OFQ-like immunoreactivity was increased significantly in some pain-modulation-related nuclei, such as ventromedial hypothalamic nucleus, raphe magnus nucleus, dorsal raphe nucleus and periaqueductal gray (PAG) after intraperitoneal (i.p.) injection of MEL 60 mg/kg, and it was further enhanced while MEL combined with EA. By using in situ hybridization, we found that ppOFQ mRNA expression was decreased in the same nuclei after the administration of MEL, and further decreased following the combination of EA and MEL. The results suggested that attenuating the release and synthesis of OFQ in the brain is one of the mechanisms that melatonin promotes acupuncture analgesia.

[Regulatory effect of tiaoxin recipe drug serum on animal's Alzheimer disease related tau protein phosphorylation].

OBJECTIVE: To study the regulatory effect and possible mechanism of Tiaoxin Recipe (TXR) on animal's Alzheimer disease related tau protein phosphorylation. METHODS: NG108 cell model was treated with Okadaic acid and related parameters were determined using MTT staining, immunoblot, coimmunoprecipitation assay, etc. RESULTS: Shown by MTT staining, NG108 cell activity decreased significantly after treated with Okadaic acid for 12 hrs, which could be ameliorated by TXR rat serum. Revealed by immunoblot method, the Okadaic acid induced elevation of phosphorylated tau protein could partly be reversed after co-treated with TXR rat serum. TXR extract could inhibit the binding of tau protein with presenilin-1, which may regulate the tau protein phosphorylation, and could be observed by coimmunoprocipitation. CONCLUSION: TXR could inhibit tau protein hyperphosphorylation, which might partially be due to the TXR caused binding of presenilin-1 with tau protein.

[Modulatory effect of orphanin FQ on the IL-1beta transcripts in hippocampus].

Using in situ hybridization and immuno-fluorescent double labeling method, we investigated the function of orphanin FQ on interlenkin-lbeta (IL-lbeta) transcripts in hippocampus. Intracerebroventricularly (icv) administrated IL-lbeta antibody reduced IL-l and TNF-alpha secreted from peritoneal macrophage. The high level of IL-lbeta transcript in hippocampus elicited by traumatic stress was blocked by icv injection of orphanin FQ (0.55 nmol), which was reversed by orphanin FQ receptor antagonist ([phe(1)Psi(CH(2)-NH)Gly(2)] nociceptin-(1-l3)-NH(2)). Opioid receptor-like receptor transcripts were found in neuron, astrocyte and microglia. Based on the results, we conclude that orphanin FQ functions as a neuroimmune modulator, and provokes immune response via mediation of IL-lbeta derived from neuron, astrocyte and microglia in hippocampus.

Systemic infusion of naloxone reduces degeneration of rat substantia nigral dopaminergic neurons induced by intranigral injection of lipopolysaccharide.

A massive degeneration of dopamine-containing neurons in the substantia nigra (SN) in the midbrain is characteristic of Parkinson's disease. Inflammation in the brain has long been speculated to play a role in the pathogenesis of this neurological disorder. Recently, we reported that treatment of primary rat mesencephalic mixed neuron-glia cultures with lipopolysaccharide (LPS) led to the activation of microglia, resident immune cells of the brain, and subsequent death of dopaminergic neurons. The LPS-induced degeneration of dopaminergic neurons was significantly attenuated by the opiate receptor antagonist (-)-naloxone and its inactive isomer (+)-naloxone, with equal potency, through an inhibition of microglial activation and their production of neurotoxic factors. In this study, injection of LPS into the rat SN led to the activation of microglia and degeneration of dopaminergic neurons: microglial activation was observed as early as 6 h and loss of dopaminergic neurons was detected 3 days after the LPS injection. Furthermore, the LPS-induced loss of dopaminergic neurons in the SN was time- and LPS concentration-dependent. Systemic infusion of either (-)-naloxone or (+)-naloxone inhibited the LPS-induced activation of microglia and significantly reduced the LPS-induced loss of dopaminergic neurons in the SN. These in vivo results combined with our cell culture observations confirmed that naloxone protects dopaminergic neurons against inflammation-mediated degeneration through inhibition of microglial activation and suggest that naloxone would have therapeutic efficacy in the treatment of inflammation-related neurological disorders. In addition, the inflammation-mediated degeneration of dopaminergic neurons in the rat SN resulting from the targeted injection of LPS may serve as a useful model to gain further insights into the pathogenesis of Parkinson's disease.

The analgesic effects of peripheral and central administration of melatonin in rats.

To explore the site and mechanism of the analgesic action of melatonin, the present study was designed to evaluate the analgesic effects of intraperitoneal (i.p.) and intracerebroventricular (i.c.v. ) administration of melatonin, and to investigate the effect of i.c. v. naloxone on the analgesic effect induced by i.p. melatonin in rats. Antinociception was determined by tail-flick latency to hot water at 50 degrees C. On i.p. administration, melatonin (30, 60 and 120 mg/kg) produced the antinociceptive effect in a dose-dependent manner, with an A(50) of 72.8 mg/kg. Administered i.c.v., melatonin (0.25, 0.5 and 1 mg/kg) also resulted in dose-dependent antinociception, with an A(50) of only 0.693 mg/kg. Injected i.c.v. to rats, 10 microg of naloxone antagonized significantly the antinociceptive effect induced by i.p. melatonin. It is concluded that melatonin has an analgesic effect in rats and the central nervous system (CNS) may be the primary site for melatonin to elicit the response, and the effect of melatonin is related to the central opioid system.

Effect of intrathecal morphine and electro-acupuncture on cellular immune function of rats and increment of mu-opioid receptor mRNA expression in PAG following intrathecal morphine.

The present study was to investigate the dynamic changes of cellular immune function of rats with intrathecal injection of (ith) morphine and the regulatory effect of electroacupuncture(EA) stimulation on "Zusanli" (St.36) and "Lanwei" (Extra 37) points. The results showed that ConA-induced rat spleen lymphocyte proliferation was significantly decreased on 2h, 4h, 8h, 12h, 24h, 48h after ith morphine(40microg/50microL). The proliferative response was recovered to nearly normal on 72h. EA on corresponding periods could prevent the decrease of lymphocyte proliferative response of rats induced by ith morphine. The same tendency was observed on the induction of IL-2 production. Further study continued to explore the mechanism of the potentiating effect of mu-opioid receptor in periaqueductal gray (PAG) and hippocampus on the immunosuppression induced by ith morphine at molecular level with in situ hybridization histochemistry technique. The results showed that ith morphine could increase the expression of mu-opioid receptor mRNA.

Effect of morphine and electro-acupuncture (EA) on apoptosis of thymocytes.

The influence of morphine and EA on the apoptosis of thymocytes were studied to investigate the posibility of its involvement in the mechanism of morphine-induced immunosuppression and the regulatory effect of EA on it. 1h after injecting 50 mg/kg morphine subcutaneously into 3-wk old Balb/c mice continually twice a day for 5 days, thymus was collected and the apoptotic cell was detected by a method of terminal deoxynucleotidyl transferase-meditaed dUTP nick end-labeling(TUNEL). The results showed that morphine significantly enhanced the percentage of TUNEL positive cells inside thymus with an appearing of apoptotic DNA ladder after 24 h incubation. Treating mice with EA of "Zusanli(St.36)" and "Lanwei(Ext.33)" for 1h after morphine administration decreased the percentage of TUNEL positive cells. EA also showed an regulatory effect on the increased the expression of CPP32 and decreased the expression of Bcl-2 by morphine. The significant enhancement of hypothalamic CRF and plasma ACTH level by morphine and the antagonize effect of EA on it suggested a possible role of Hypothalamus-pituitary-adrenal (HPA) axis played in the apoptosis of thymocytes by morphine and the regulatory effect of EA.

Selective MT(2) melatonin receptor antagonist blocks melatonin-induced antinociception in rats.

The present study was undertaken to assess the effects of intracerebroventricular (i.c.v.) luzindole (a selective MT(2) melatonin receptor antagonist) and prazosin (a selective MT(3) melatonin receptor antagonist) on melatonin-induced antinociception, so as to clarify which of melatonin receptor subtypes within the central nervous system (CNS) was mediating antinociception. The pain threshold of rats was measured by the hot water (50 degrees C) tail-flick test. It was found that intraperitoneal (i.p.) melatonin (30, 60, 120 mg/kg) resulted in a dose-dependent antinociception. Luzindole (50, 100 microgram) administered intracerebroventricularly antagonized significantly the antinociceptive effect induced by i.p. melatonin (120 mg/kg), whereas prazosin (50 microgram) did not. Neither luzindole (100 microgram, i.c.v.) nor prazosin (50 microgram, i.c.v.) affected the nociceptive threshold when given alone. The results suggest that melatonin-induced antinociception is mediated through the MT(2) melatonin receptor subtype within the CNS.

Role of dopamine receptors and the changes of the tyrosine hydroxylase mRNA in acupuncture analgesia in rats.

Previous studies have shown that dopamine (DA) is involved in electroacupuncture analgesia (EAA). L-tetrahydropalmatine (l-THP), a DA receptor antagonist was proved to potentiate EAA in both laboratory research and clinical practice. In the present study SK&F-38393 and quinpirole (Qui), selective agonists of D1 or D2 receptors respectively were injected into nucleus (N.) accumbens of rats to investigate the roles of D1 and D2 receptors in the potentiation of EAA induced by l-THP. The injection of D1 agonist SK&F-38393 (5 microg or 10 microg) attenuated the potentiation of EAA induced by l-THP, 10 microg SK&F-38393 attenuated EAA as well, while the injection of D2 agonist Qui (10 microg or 20 microg) had no effect on EAA and the potentiation of EAA induced by l-THP. DA release was shown to increase in EAA in previous work, however, whether the synthesis of DA was influenced is still unknown. In the present study, dot blot technique was applied to observe the effect of noxious stimulation or electroacupuncture on the level of tyrosine hydroxylase (TH) mRNA in rat brain. Noxious electric stimulation was found to elevate the TH mRNA level in substantia nigra (SN) and hypothalamus, while electro-acupuncture attenuated the effect of noxious stimulation on TH mRNA. The results indicate that D1 but not D2 receptor in N. accumbens plays an important role in EAA. EA might regulate the biosynthesis of DA by altering the TH gene transcription.

Supraspinal hyperalgesia and spinal analgesia by [Phe1psi(CH2-NH)Gly2]nociceptin-(1-13)-NH2 in rat.

[Phe1psi(CH2-NH)Gly2]nociceptin-(1-13)-NH2, a pseudopeptide analog of nociceptin, was originally seen as an antagonist of nociceptin receptors. In the present study, it was observed that intracerebroventricular (i.c.v.) injection of this pseudopeptide (1, 5, 10 microg) significantly decreased the tail-flick latency of rats, indicating a hyperalgesic effect, while intrathecal (i.t.) injection of it (1, 2.5, 10 microg) dramatically increased the tail-flick latency, indicating an analgesic effect. This strengthened the in vivo evidence that [Phe1psi(CH2NH)Gly2]nociceptin-(1-13)-NH2 might be an agonist of nociceptin receptors.

Effects of orphanin FQ on endomorphin-1 induced analgesia.

Orphanin FQ (also known as nociceptin) is a 17-amino-acid peptide which acts as a potent endogenous agonist of the orphan opioid receptor-like (ORL1) receptor. Endomorphin-1, a 4-amino-acid peptide discovered recently, is a potent and selective endogenous agonist for the mu-opiate receptor. In the present study, the effect of OFQ or/and endomorphin-1 on the response to noxious thermal stimuli was observed using the tail-flick test in rats. Intracerebroventricular (i.c.v.) administration of OFQ (1, 5 microg) could shorten tail-flick latency; In contrast, intrathecal (i.t.) administration of OFQ (1, 2 or 10 microg) could increase the latency; i.c.v. (1, 2, 5 microg) or i.t. (0.2, 2, 5 microg) administration of endomorphin-1 dose-dependently increased the latency, indicating an analgesic effect. Furthermore, OFQ (0.1-5 microg) when intraventricularly injected together with endomorphin-1 (5 microg), could dose-dependently reverse the analgesia induced by the latter. On the contrary, OFQ (1 microg) intrathecally injected together with endomorphin-1 (0.2 microg) could further increase the tail-flick latency. The results showed that OFQ at the supraspinal level produces hyperalgesia and is antagonistic to endomorphin-1, while at the spinal level it produces analgesia and is synergic with endomorphin-1. Different interaction mechanism between OFQ and endomorphin-1 in the brain and the spinal cord is thus suggested.