Descending serotonergic modulation from rostral ventromedial medulla to spinal trigeminal nucleus is involved in experimental occlusal interference-induced chronic orofacial hyperalgesia.

BACKGROUND: Dental treatment associated with unadaptable occlusal alteration can cause chronic primary myofascial orofacial pain. The serotonin (5-HT) pathway from the rostral ventromedial medulla (RVM) exerts descending modulation on nociceptive transmission in the spinal trigeminal nucleus (Sp5) and facilitates chronic pain. The aim of this study was to investigate whether descending 5-HT modulation from the RVM to the Sp5 is involved in the maintenance of primary myofascial orofacial hyperalgesia after persistent experimental occlusal interference (PEOI) or after delayed removal of experimental occlusal interference (REOI). METHODS: Expressions of 5-HT3A and 5-HT3B receptor subtypes in the Sp5 were assessed by immunofluorescence staining and Western blotting. The release and metabolism of 5-HT in the Sp5 were measured by high-performance liquid chromatography. Changes in the pain behavior of these rats were examined after specific pharmacologic antagonism of the 5-HT3 receptor, chemogenetic manipulation of the RVM 5-HT neurons, or selective down-regulation of 5-HT synthesis in the RVM. RESULTS: Upregulation of the 5-HT3B receptor subtype in the Sp5 was found in REOI and PEOI rats. The concentration of 5-HT in Sp5 increased significantly only in REOI rats. Intrathecal administration of Y-25130 (a selective 5-HT3 receptor antagonist) dose-dependently reversed the hyperalgesia in REOI rats but only transiently reversed the hyperalgesia in PEOI rats. Chemogenetic inhibition of the RVM 5-HT neurons reversed the hyperalgesia in REOI rats; selective down-regulation of 5-HT in advance also prevented the development of hyperalgesia in REOI rats; the above two manipulations did not affect the hyperalgesia in PEOI rats. However, chemogenetic activation of the RVM 5-HT neurons exacerbated the hyperalgesia both in REOI and PEOI rats. CONCLUSIONS: These results provide several lines of evidence that the descending pathway from 5-HT neurons in the RVM to 5-HT3 receptors in the Sp5, plays an important role in facilitating the maintained orofacial hyperalgesia after delayed EOI removal, but has a limited role in that after persistent EOI.

MTUS1 is a promising diagnostic and prognostic biomarker for colorectal cancer.

BACKGROUND: The morbidity and mortality of colorectal cancer (CRC) remain high, posing a serious threat to human life and health. The early diagnosis and prognostic evaluation of CRC are two major challenges in clinical practice. MTUS1 is considered a tumour suppressor and can play an important role in inhibiting cell proliferation, migration, and tumour growth. Moreover, the expression of MTUS1 is decreased in different human cancers, including CRC. However, the biological functions and molecular mechanisms of MTUS1 in CRC remain unclear. METHODS: In the present study, data from The Cancer Genome Atlas (TCGA) database were analysed using R statistical software (version 3.6.3.) to evaluate the expression of MTUS1 in tumour tissues and adjacent normal tissues using public databases such as the TIMER and Oncomine databases. Then, 38 clinical samples were collected, and qPCR was performed to verify MTUS1 expression. We also investigated the relationship between MTUS1 expression and clinicopathological characteristics and elucidated the diagnostic and prognostic value of MTUS1 in CRC. In addition, the correlation between MTUS1 expression and immune infiltration levels was identified using the TIMER and GEPIA databases. Furthermore, we constructed and analysed a PPI network and coexpression modules of MTUS1 to explore its molecular functions and mechanisms. RESULTS: CRC tissues exhibited lower levels of MTUS1 than normal tissues. The logistic regression analysis indicated that the expression of MTUS1 was associated with N stage, TNM stage, and neoplasm type. Moreover, CRC patients with low MTUS1 expression had poor overall survival (OS). Multivariate analysis revealed that the downregulation of MTUS1 was an independent prognostic factor and was correlated with poor OS in CRC patients. MTUS1 expression had good diagnostic value based on ROC analysis. Furthermore, we identified a group of potential MTUS1-interacting proteins and coexpressed genes. GO and KEGG enrichment analyses showed that MTUS1 was involved in multiple cancer-related signalling pathways. Moreover, the expression of MTUS1 was significantly related to the infiltration levels of multiple cells. Finally, MTUS1 expression was strongly correlated with various immune marker sets. CONCLUSIONS: Our results indicated that MTUS1 is a promising biomarker for predicting the diagnosis and prognosis of CRC patients. MTUS1 can also become a new molecular target for tumour immunotherapy.

Astrocytes in the rostral ventromedial medulla contribute to the maintenance of oro-facial hyperalgesia induced by late removal of dental occlusal interference.

BACKGROUND: Astrocytes in the rostral ventromedial medulla (RVM) contribute to descending pain modulation, but their role in oro-facial pain induced by persistent experimental dental occlusal interference (PEOI) or following EOI removal (REOI) is unknown. OBJECTIVE: To explore the involvement of RVM astrocytes in PEOI-induced oro-facial hyperalgesia or its maintenance following REOI. METHODS: Male rats were randomly assigned into five groups: sham-EOI, postoperative day 6 and 14 of PEOI (PEOI 6 d and PEOI 14 d), postoperative day 6 following REOI on day 3 (REOI 3 d) and postoperative day 14 following REOI on day 8 (REOI 8 d). The nociceptive head withdrawal threshold (HWT) and activities of RVM ON- or OFF-cells were recorded before and after intra-RVM astrocyte gap junction blocker carbenoxolone (CBX) microinjection. RVM astrocytes were labelled immunohistochemically with glial fibrillary acidic protein (GFAP) and analysed semi-quantitatively. RESULTS: Persistent experimental dental occlusal interference-induced oro-facial hyperalgesia, as reflected in decreased HWTs, was partially inhibited by REOI at day 3 but not at day 8 after EOI placement. Increased GFAP-staining area occurred only in REOI 8 d group in which CBX could inhibit the maintained hyperalgesia; CBX was ineffective in inhibiting hyperalgesia in PEOI 14 d group. OFF-cell activities showed no change, but the spontaneous activity and responses of ON-cells were significantly enhanced that could be suppressed by CBX in REOI 8 d group. CONCLUSION: Rostral ventromedial medulla astrocytes may not participate in PEOI-induced oro-facial hyperalgesia or hyperalgesia inhibition by early REOI but are involved in the maintenance of oro-facial hyperalgesia by late REOI.

Influence of lymph node involvement or lymphadenectomy on prognosis of patients with borderline ovarian tumors: A systematic review and meta-analysis.

OBJECTIVE: Borderline ovarian tumors (BOTs) account for about 15% of all epithelial tumors of the ovary, and around 75% of patients are diagnosed in early stages. Although many of these patients have lymph node involvement (LNI), whether LNI decreases their survival is controversial, raising the question of whether lymphadenectomy should be performed. We conducted a systematic review and meta-analysis of these questions. METHODS: We searched articles related to LNI and lymphadenectomy in patients with BOTs in PubMed, EMBASE, and the Cochrane Central Register of Controlled Trials. Data on rate of LNI, recurrence and survival were pooled and meta-analyzed using a random-effects model. Heterogeneity was evaluated using the I2 test. RESULTS: A total of 25 studies with 12,503 patients were meta-analyzed. The overall pooled rate of LNI was 10% [95% confidence interval (CI) 0.07-0.13]. LNI was associated with a higher risk of recurrence [odds ratio (OR) 2.23, 95% CI 1.13-4.40]. However, LNI did not significantly affect cause-specific survival [hazard ratio (HR) 1.73, 95% CI 0.99-3.02] or disease-free survival (HR 1.48, 95% CI 0.56-3.92). Similarly, lymphadenectomy did not significantly affect risk of recurrence (OR 0.91, 95% CI 0.57-1.46), overall survival (HR 0.90, 95% CI 0.58-1.40), disease-free survival (HR 0.95, 95% CI 0.61-1.50) or progression-free survival (HR 0.60, 95% CI 0.24-1.49). CONCLUSIONS: LNI appears to increase risk of recurrence in BOT patients, but neither it nor lymphadenectomy appears to influence prognosis. Therefore, lymphadenectomy should be considered only for certain BOT patients, such as those with suspected LNI based on imaging or surgical exploration.

17ß-Estradiol Exacerbated Experimental Occlusal Interference-Induced Chronic Masseter Hyperalgesia by Increasing the Neuronal Excitability and TRPV1 Function of Trigeminal Ganglion in Ovariectomized Rats.

Pain symptoms in temporomandibular disorders (TMD) predominantly affect reproductive women, suggesting that estrogen regulates pain perception. However, how estrogen contributes to chronic TMD pain remains largely unclear. In the present study, we performed behavioral tests, electrophysiology, Western blot and immunofluorescence to investigate the role and underlying mechanisms of estrogen in dental experimental occlusal interference (EOI)-induced chronic masseter mechanical hyperalgesia in rats. We found that long-term 17ß-estradiol (E2) replacement exacerbated EOI-induced masseter hyperalgesia in a dose-dependent manner in ovariectomized (OVX) rats. Whole-cell patch-clamp recordings demonstrated that E2 (100 nM) treatment enhanced the excitability of isolated trigeminal ganglion (TG) neurons in OVX and OVX EOI rats, and EOI increased the functional expression of transient receptor potential vanilloid-1 (TRPV1). In addition, E2 replacement upregulated the protein expression of TRPV1 in EOI-treated OVX rats. Importantly, intraganglionic administration of the TRPV1 antagonist AMG-9810 strongly attenuated the facilitatory effect of E2 on EOI-induced masseter mechanical sensitivity. These results demonstrate that E2 exacerbated EOI-induced chronic masseter mechanical hyperalgesia by increasing TG neuronal excitability and TRPV1 function. Our study helps to elucidate the E2 actions in chronic myogenic TMD pain and may provide new therapeutic targets for relieving estrogen-sensitive pain.

The relationship between frequency dependence of action potential duration and the expression of TRPC3 in rabbit ventricular myocardium.

BACKGROUND/AIMS: Cardiac action potential duration (APD) is regulated by heart rate, leading to the trans-membrane movement of inorganic ions. Whether the alteration of heart rate can affect the expression of transient receptor potential canonical channels (TRPCs), further studies should be made. We investigated the changes of APD at different stimulus frequencies and their influences on the expression of TRPCs in rabbit ventricular myocardium. METHODS: Monophasic action potential (MAP) was recorded by contact electrode technique in different programmed stimulus frequencies on rabbit ventricular epicardium in vivo, and the expression of TRPCs was detected using RT-PCR and Western blot. RESULTS: At the frequency range of 4.5-7.5 Hz, APD gradually shortened with the increase of stimulus frequency, showing the property of significant frequency dependence in rabbit ventricular myocardium in vivo. Compared with 4.5 Hz group, TRPC3 mRNA and protein expression increased in 6 Hz and 7.5 Hz groups by way of frequency dependence. Both amiodarone (AM) and neferine (Nef) could prolongate APD and showed characters of frequency independence at the designed frequency. In contrast with 4.5 Hz control group, it was Nef treatment group rather than AM treatment group that could obviously increase the expression of TRPC3 mRNA and protein. CONCLUSIONS: At the frequency range of 4.5-7.5 Hz, frequency-dependent shortening of APD was associated with the expression of TRPC3. AM and Nef exhibited frequency-independent lengthening of the APD. Nef may prolong APD via the increasement of TRPC3.

Upregulation of TRPC1 contributes to contractile function in isoproterenol-induced hypertrophic myocardium of rat.

AIMS: The transient receptor potential canonical channel 1 (TRPC1) is a crucial component of the stretch-activated ion channels (SACs). The objective of this research was to demonstrate the contribution of TRPC1 in maintaining cardiac contractile function in the hypertrophic myocardium. METHODS: Hypertrophic rat hearts were induced by injecting isoproterenol intraperitoneally, and the expressions of TRPC1/3/6 and Na(+)/Ca(2+) exchanger 1 (NCX1) proteins were analyzed by Western blot. The intracellular calcium images, the action potential of myocardium, the length-dependent contractile force of ventricle muscle and the cardiac output of isolated heart were investigated. RESULTS: The expression of TRPC1 was increased in the hypertrophic myocardium. After being stretched, the ascendant amplitude of the increase in the intracellular calcium ion concentration ([Ca(2+)]i) in the hypertrophic myocardium was higher than that in the normal myocardium. The increase of the APD50 and the amplitude of the membrane potential depolarization were more significant in the hypertrophic myocardium after the activation of SACs. When the heart preparations were perfused with Tyrode's solution, there was no difference in the cardiac systolic function between the cardiac hypertrophy group and the control group. Gadolinium, a SACs blocker, reduced the length-dependent contractile force and suppressed the ascending limb of the Frank-Starling curves in the hypertrophic heart. CONCLUSIONS: The upregulation of TRPC1 contributes to the contractile function in the hypertrophic myocardium by increasing [Ca(2+)]i through the SACs.

The formation mechanism of the bilayer capsular contracture after an augmentation mammoplasty with a rough-surface prosthesis and its prevention and treatment.

BACKGROUND: The present study aimed to explore the formation mechanism of the bilayer capsular contracture after augmentation mammoplasty with a rough-surface prosthesis and its prevention and treatment. METHODS: The nursing process, clinical signs, intraoperative findings, and pathological data after an augmentation mammoplasty with rough-surface prosthesis were observed and collected, the formation mechanism of the bilayer capsular contracture was analyzed, and the prevention and treatment were also discussed. RESULTS: A total of 18 patients were included into the present study, among which 15 patients underwent capsule relaxation plus secondary augmentation mammoplasty and three patients encountered a single-layer capsular contracture after the operation; the recurrence rate was 16%. All patients were followed up for 1-13 years without a presentation of recurrence. CONCLUSIONS: The formation of the bilayer capsular contracture after augmentation mammoplasty is correlated with the formation of the inner capsule, inadequate separation of cavities, foreign body reaction, and an improper massage of the breasts, and the effective preventive measures include removing new cavities, resecting the capsular contracture capsule, stopping bleeding, replanting a rough-surface or smooth prosthesis, and correcting breast massaging.

Proteomics and liquid biopsy characterization of human EMT-related metastasis in colorectal cancer.

Tumor cells undergo epithelial-mesenchymal transition (EMT), however, there is a room of disagreement in role of EMT heterogeneity to colorectal cancer metastasis (mCRC) evolution. To uncover new EMT-related metastasis proteins and pathways, we addressed the EMT status in colorectal cancer liver metastasis patient-derived CTCs to identify proteins that promote their distant metastasis. And then, we performed a comparative proteomic analysis in matched pairs of primary tumor tissues, adjacent mucosa tissues and liver metastatic tissues. By integrative analysis we show that, unstable Epithelial/Mesenchymal (E/M)-type CTCs had the strongest liver metastases formation ability and the proportion of E/M-type CTCs correlated with distant metastases. Using an optimized proteomic workflow including data independent acquisition (DIA) and parallel reaction monitoring (PRM), we identified novel EMT-related protein cluster (GNG2, COL6A1, COL6A2, DCN, COL6A3, LAMB2, TNXB, CAVIN1) and well-described (ERBB2) core protein level changes in EMT-related metastasis progression, and the proteomic data indicate ERBB2, COL6A1 and CAVIN1 are promising EMT-related metastatic biomarker candidates. This study contributes to our understanding of the role that EMT plays in CRC metastasis and identifies heterogeneous EMT phenotypes as a key piece for tumor progression and prognosis. We further propose that therapies targeting this aggressive subset (E/M-type) of CTCs and related protein may be worthy of exploration as potential suppressors of metastatic evolution.

OX40 as a novel target for the reversal of immune escape in colorectal cancer.

First-generation immunological checkpoint inhibitors, such as CTLA-4, PD-L1 and PD-1 exhibit significant advantages over conventional cytotoxic drugs, such as oxaliplatin and 5-FU, for the treatment of colorectal cancer. However, these inhibitors are not ideal due to their low objective response rate and the vulnerability of these treatment methods when faced with emerging drug resistant cancers. This study summarizes the immunological characteristics of colorectal cancer treatment, and analyzes the ways in which OX40 may improve the efficacy of these treatments. Activation of the OX40 signaling pathway can enhance the activity of CD4+/CD8+ T cells and inhibit the function of Treg. Simultaneously, OX40 can directly inhibit the expression of Foxp3, affect the inhibitory function of Treg, and inhibit the immunosuppressive factors in the tumor microenvironment so as to reverse immune escape and reverse drug resistance. Therefore, OX40 is an important target for treating colorectal cancer in "cold tumors" with less immunogenicity.

Anlotinib suppresses metastasis and multidrug resistance via dual blockade of MET/ABCB1 in colorectal carcinoma cells.

Anlotinib, a highly selective multi-targeted tyrosine kinase inhibitor (TKI) has therapeutic effects on non-small-cell lung cancer (NSCLC). In this study, the anti-tumor activity and molecular mechanism of anlotinib in metastatic colorectal cancer (mCRC) was explored. The anti-angiogenesis, anti-metastasis, anti-proliferative, and anti-multidrug resistance efficacy of anlotinib were analyzed by using in vitro and in vivo models of human CRC cells. The results indicated that anlotinib boosted chemo-sensitivity of CRC cells, and restrained its proliferation. Besides the suppression of the MET signaling pathway, anlotinib also inhibited invasion and migration of CRC cells. Furthermore, anlotinib prevented VEGF-induced angiogenesis, N-cadherin (CDH2)-induced cell migration, and reversed ATP-binding cassette subfamily B member 1 (ABCB1) -mediated CRC multidrug resistance in CRC. The CRC liver metastasis and subcutaneously implanted xenograft model testified that anlotinib could inhibit proliferation and liver metastasis in CRC cells. Such an observation suggested that a combination of anlotinib with anti-cancer drugs could attenuate angiogenesis, metastasis, proliferative, and multidrug resistance, which constitutes a novel treatment strategy for CRC patients with metastasis.

The microfloral analysis of secondary caries biofilm around Class I and Class II composite and amalgam fillings.

BACKGROUND: Secondary caries is responsible for 60 percent of all replacement restorations in the typical dental practice. The diversity of the bacterial sources and the different types of filling materials could play a role in secondary caries. The aim of this study was to determine and compare the microbial spectrum of secondary caries biofilms around amalgam and composite resin restorations. METHODS: Clinical samples were collected from freshly extracted teeth diagnosed with clinical secondary caries. Samples were categorized into four groups according to the types of restoration materials and the classification of the cavity. Biofilms were harvested from the tooth-restoration interface using a dental explorer and after dilution were incubated on special agars. The bacteria were identified using the biochemical appraisal system. Statistical calculations were carried out using SPSS11.5 software to analyze the prevalence of the bacteria involved in secondary caries. RESULTS: Samples from a total of four groups were collected: two groups were collected from amalgam restorations, each had 21 samples from both Class I and Class II caries; and the other two groups were from composite resin restorations, each had 13 samples from both class I and class II caries. Our results showed: (1) Anaerobic species were dominant in both restoration materials. (2) In terms of the types of individual bacteria, no significant differences were found among the four groups according to the geometric mean of the detected bacteria (P > 0.05). However, there were significant differences among the detected bacteria within each group (P < 0.05). The composition of each bacterium had no statistical difference among the four groups (P > 0.05), but showed significant differences among the detected bacteria in each group (P < 0.05). (3) Among the four groups, there were no significant differences for the detection rate of each bacterium (P > 0.05), however, the detection rate of each bacterium within each group was statistically different among the detected bacteria (P < 0.05). CONCLUSIONS: The proportion of obligatory anaerobic species was much greater than the facultative anaerobic species in the biofilm of secondary caries. Statistically, the materials of restoration and the location of secondary caries did not show any significant effects on the composition of the microflora.

The PacBio Full-Length Transcriptome of the Tea Aphid as a Reference Resource.

The tea aphid, Aphis aurantii, has become one of the destructive pests in tea plantations in the tropics and subtropics. Very few functional studies have so far focused on the developmental and reproductive biology at a molecular level, because of the lack of comprehensive genetic information. Full-length transcriptomes represent a very highly efficient approach to obtain reference gene sequences in non-model insects. In the present study, the transcriptome of A. aurantii was comprehensively sequenced using PacBio Iso-Seq technology. A total of 46.8 Gb nucleotides and 15,938 non-redundant full-length transcripts were obtained, 13,498 (84.69%) of which were annotated into seven databases. Of these transcripts, 2,029 alternative splicing events and 15,223 simple sequence repeats were detected. Among these transcripts, 4,571 (28.68%) and 11,367 (71.32%) were long non-coding RNAs (lncRNAs) and protein-coding genes, respectively. Five hundred and ninety transcription factors were detected. The first full-length transcriptome represents a significant increase in the known genetic information of A. aurantii. It will assist the future functional study of genes involved in its development and reproduction.

Ovarian metastasis in women with cervical carcinoma in stages IA to IIB: A systematic review and meta-analysis.

BACKGROUND: Cervical cancer is one of the common malignancies that afflict women worldwide. In rare cases, cervical cancer leads to ovarian metastasis (OM), resulting in poor outcomes. We conducted a systematic review and meta-analysis to evaluate the incidence and risk factors of OM in patients with adenocarcinoma (ADC) or squamous cell carcinoma (SCC) of the cervix. METHODS: We searched articles focused on OM in cervical carcinoma in PubMed, Embase, and the Cochrane Central Register of Controlled Trials. A meta-analysis was performed including selected publications. Pooled odds ratio (OR) and 95% confidence interval (95% CI) were calculated using random-effects models. The heterogeneity was evaluated by the I test. I > 50% was considered high heterogeneity. RESULTS: A total of 12 studies with 18,389 patients with cervical cancer in International Federation of Gynecology and Obstetrics stages IA to IIB were included in the meta-analysis. The overall incidence of OM was 3.61% among patients with ADC and 1.46% among patients with SCC (ADC vs SCC: OR 3.89, 95% CI 2.62-5.78; P < .001). Risk factors for OM were age >40 years (OR 1.79, 95% CI 1.02-3.13), bulky tumor (OR 2.65, 95% CI 1.77-3.95), pelvic lymph node involvement (PLNI; OR 9.33, 95% CI 6.34-13.73), lymphovascular space involvement (LVSI; OR 4.38, 95% CI 1.86-10.31), parametrial invasion (PMI; OR 7.87, 95% CI 5.01-12.36), and corpus uteri invasion (CUI; OR 7.64, 95% CI 2.51-23.24). PLNI, LVSI, and PMI were the leading risk factors, contributing to OM with respective population attributable fractions of 64.8%, 58.8%, and 51.5%. CONCLUSION: The incidence of OM is relatively low in ADC and SCC patients. Risk factors for OM include PLNI, LVSI, PMI, bulky tumor, CUI, or age over 40 years, with the first 3 contributing more to risk of OM.

Contraceptive and reproductive health practices of unmarried women globally, 1999 to 2018: Systematic review and meta-analysis.

BACKGROUND: Premarital sex practices and contraceptive prevalence rate (CPR) among unmarried women worldwide remain unclear, even though unmarried women tend to have multiple sex partners over time, which makes their sexual behaviors particularly important to the sexual and reproductive health of society more broadly. METHODS: We searched the MEDLINE, PubMed, and Google Scholar databases for relevant articles published between January 1, 1999 and December 31, 2018. Data on prevalence of premarital sexual intercourse, use of highly prevalent contraceptive methods, as well as CPR overall and at first sexual intercourse were extracted and estimated using a DerSimonian- Laird random effects model. RESULTS: Of the 3918 articles identified, 37 covering 19 countries were included. The estimated overall prevalence of premarital sexual intercourse was 41.9% (95%CI 34.2-49.6%). Pooled CPR was 57.0% (95%CI 44.3-69.8%) overall and 57.6% (95% CI 39.5- 75.6%) at first intercourse. The overall prevalence of condom use was 51.2% (95%CI 42.7-59.7%), followed by oral contraceptives (20.5%, 95%CI 13.7-27.3%), withdrawal (12.7%, 95%CI 9.4-15.9%), and rhythm (12.1%, 95%CI 6.7-17.4%). CONCLUSION: The findings of this global study indicate worrying trends in unprotected intercourse and contraceptive practices, suggesting the need for greater attention and resources aimed at educating unmarried adolescent women about sexual and reproductive health. SYSTEMATIC REVIEW REGISTRATION NUMBER: CRD42019132736.

Genistein Antagonizes 17ß-Estradiol Effects on Glutamate-Evoked Masseter Muscle Hypernociception in Rats.

Temporomandibular disorders (TMDs) predominantly affect women of reproductive ages, with pain as the main symptom. The aim of the present study was to examine the effects of 17ß-estradiol (E2) on glutamate-evoked hypernociception of masseter muscle and to examine whether genistein could antagonize the effects of E2 in female rats. Injection of glutamate into the masseter muscle dose-dependently decreased head withdrawal thresholds, a parameter for mechanical hypernociception. Head withdrawal thresholds in ovariectomized rats also decreased with increasing doses of E2 replacement, and were further aggravated by injection of glutamate (1M, 40µL) into the masseters. Genistein at doses of 7.5 and 15 mg/kg antagonized E2-induced hypernociception of masseter muscle, and at doses of 7.5, 15, and 30 mg/kg also antagonized E2 potentiation of glutamate-evoked hypernociception of masseter muscle. Genistein produced optimal antagonistic effects of E2 on nociception behavior at a dose of 15 mg/kg. On the molecular level, tyrosine phosphorylation of the NR2B subunit of the N-methyl-D-aspartate receptor (pNR2B) and phosphorylated mitogen-activated protein kinase (pERK1/2) were significantly upregulated in the hippocampus following glutamate injection and were further potentiated by E2 replacement. Genistein at dose of 15 mg/kg partially reversed E2-potentiated glutamate-evoked upregulation of pNR2B and pERK1/2 expression in the hippocampus. These results indicated that moderate doses of genistein could antagonize E2 enhanced glutamate-evoked hypernociception of masseter muscle possibly via N-methyl-D-aspartate receptor and ERK1/2 signaling pathways in the hippocampus.

Na (+) /Ca (2+) Exchanger 3 is Downregulated in the Hippocampus and Cerebrocortex of Rats with Hyperthermia-induced Convulsion.

BACKGROUND: Na + /Ca 2+ exchanger (NCX) plays a crucial role in pentylenetetrazol-induced convulsion. However, it is unclear whether NCX is critically involved in hyperthermia-induced convulsion. In this study, we examined the potential changes in NCX3 in the hippocampus and cerebrocortex of rats with hyperthermia-induced convulsion. METHODS: Twenty-one Sprague Dawley rats were randomly assigned to control group, convulsion-prone group and convulsion-resistant group (n = 7 in each group). Whole-cell patch-clamp method was used to record NCX currents. Both the Western blotting analysis and immunofluorescence labeling techniques were used to examine the expression of NCX3. RESULTS: NCX currents were decreased in rats after febrile convulsion. Compared to the control group, NCX3 expression was decreased by about 40% and 50% in the hippocampus and cerebrocortex of convulsion-prone rats, respectively. Furthermore, the extent of reduction in NCX3 expression seemed to correlate with the number of seizures. CONCLUSIONS: There is a significant reduction in NCX3 expression in rats with febrile convulsions. Our findings also indicate a potential link between NCX3 expression, febrile convulsion in early childhood, and adult onset of epilepsy.

Anti-fibrotic effects of neferine on carbon tetrachloride-induced hepatic fibrosis in mice.

The effects of neferine, a bisbenzylisoquinline alkaloid extracted from the seed embryo of the Chinese traditional medicine Nelumbo nucifera Gaertn, on carbon tetrachloride (CCl4)-induced hepatic fibrosis in mice were evaluated. Adult male Kunming mice were administered with CCl4 1 ml/kg via intraperitoneal injection twice a week for 8 weeks. At the beginning of the 9th week, mice were treated with normal saline, colchicine (0.1 mg/kg), and neferine (5, 10, 20 mg/kg) via intraperitoneal injection once a day for 2 weeks. The liver index and histological examination, plasma ALT/AST levels, hydroxyproline and TGF-ß1 content of liver tissue were examined. In the model group, the liver index, the hydroxyproline content of liver tissue and plasma ALT/AST levels were increased, and a high expression of TGF-ß1 was observed. The abnormal changes could be improved by neferine in a dose-dependent manner. Our data showed that neferine had an antifibrosis effect on CCl4-induced hepatic fibrosis in mice, possibly partly due to the decreased expression of TGF-ß1 in the liver.

Protective effect of heme oxygenase-1 on Wistar rats with heart failure through the inhibition of inflammation and amelioration of intestinal microcirculation.

BACKGROUND: Myocardial infarction (MI) has likely contributed to the increased prevalence of heart failure (HF). As a result of reduced cardiac function, splanchnic blood flow decreases, causing ischemia in villi and damage to the intestinal barrier. The induction of heme oxygenase-1 (HO-1) could prevent, or lessen the effects of stress and inflammation. Thus, the effect and mechanism thereof of HO-1 on the intestines of rats with HF was investigated. METHODS: Male Wistar rats with heart failure through ligation of the left coronary artery were identified with an left ventricular ejection fraction of < 45% through echocardiography and then divided into various experimental groups based on the type of peritoneal injection they received [MI: saline; MI + Cobalt protoporphyrin (CoPP): CoPP solution; and MI + Tin mesoporphyrin IX dichloride (SnMP): SnMP solution]. The control group was comprised of rats without coronary ligation. Echocardiography was performed before ligation for a baseline and eight weeks after ligation in order to evaluate the cardiac function of the rats. The bacterial translocation (BT) incidence, mesenteric microcirculation, amount of endotoxins in the vein serum, ileum levels of HO-1, carbon oxide (CO), nitric oxide (NO), interleukin (IL)-10, tumour necrosis factor-α (TNF-α), and the ileum morphology were determined eight weeks after the operation. RESULTS: The rats receiving MI + CoPP injections exhibited a recovery in cardiac function, an amelioration of mesenteric microcirculation and change in morphology, a lower BT incidence, a reduction in serum and ileac NO and TNF-α levels, and an elevation in ileac HO-1, CO, and interleukin-10 (IL-10) levels compared to the MI group (P < 0.05). The rats that received the MI + SnMP injections exhibited results inverse to the MI (P < 0.05) group. CONCLUSIONS: HO-1 exerted a protective effect on the intestines of rats with HF by inhibiting the inflammation and amelioration of microcirculation through the CO pathway. This protective effect could be independent from the recovery of cardiac function.