A multicenter, randomized controlled study on the efficacy of agomelatine in ameliorating anhedonia, reduced motivation, and circadian rhythm disruptions in patients with major depressive disorder (MDD).

OBJECTIVE: To evaluate the clinical efficacy and safety of Agomelatine in improving symptoms in patients with major depressive disorder (MDD), providing more scientific evidence for the treatment of depression, and offering more effective therapeutic options for patients. METHODS: A total of 180 MDD patients in acute phase from 10 psychiatric hospitals of Grade three in Zhejiang Province were enrolled in this 12-week study with the competitive and consecutive pattern, and they were randomized into two different groups treated with flexible-dosage antidepressants of selective serotonin reuptake inhibitors (SSRI) or agomelatine, respectively. The subjects were evaluated with psychological scales of HAMD-17, HAMA, SHAPS for anhedonia, MFI-20 for fatigue, PQSI for sleep quality and MEQ for disturbances in chronobiologic rhythms at baseline, 2, 4, 8 and 12-weekend points, and TESS was used for side-effect. The results were analyzed with repeated measurement analysis of variance. RESULTS: The two groups each had 90 participants, and there were no significant differences at baseline. The scores of various assessment scales showed statistically significant time main effects during the visits (P < 0.01). The Agomelatine group demonstrated faster efficacy within 2 weeks, with better improvement in SHAPS, MEQ, and PSQI compared to the SSRIs group. However, the remission rate at 12 weeks was lower in the Agomelatine group than in the SSRIs group (63.3% and 72.2%), but the difference between the groups was not statistically significant. The Agomelatine group had fewer adverse reactions (14.4% and 16.7%), but there was a slightly higher incidence of liver function impairment (6.7% and 4.4%), with no statistically significant difference between the groups. CONCLUSION: Agomelatine, as a novel antidepressant, shows certain advantages in improving depression and anxiety symptoms and is comparable to SSRIs in terms of safety. However, its long-term efficacy and safety on MDD or other depressive subtypes still require further observation and research.

Smartphone addiction and victimization predicts sleep problems and depression among children.

BACKGROUND: In this study we examined the phenomena of smartphone addiction, online harassment, and school bullying/victimization to predict the prospective influence these could have on the onset and persistence of sleep problems and depression among children. METHODS: Responses from 2155 fifth-grade children recruited from 30 primary schools in Taipei were assessed, and a follow-up was performed in the 6th grade. Self-administered questionnaires were collected for each year. FINDINGS: Children who reported smartphone addictions, online harassment, and school bullying/victimization coupled with an increase in those factors were more likely to experience the onset and persistence of sleep problems. In addition, children who reported smartphone addiction, online harassment, school bullying/victimization, and poor sleep quality were more likely to experience the onset and persistence of depression. IMPLICATIONS: School nurses or pediatric nurses should be able to assess children's Internet use and risks to understand potential influences on sleep quality and mental status and provide recommendations for children, parents and schools.

[In-vivo and ex-vivo studies on region-specific remodeling of large elastic arteries due to simulated weightlessness and its prevention by gravity-based countermeasure].

The present study was designed to test the hypothesis that a medium-term simulated microgravity can induce region-specific remodeling in large elastic arteries with their innermost smooth muscle (SM) layers being most profoundly affected. The second purpose was to examine whether these changes can be prevented by a simulated intermittent artificial gravity (IAG). The third purpose was to elucidate whether vascular local renin-angiotensin system (L-RAS) plays an important role in the regional vascular remodeling and its prevention by the gravity-based countermeasure. This study consisted of two interconnected series of in-vivo and ex-vivo experiments. In the in-vivo experiments, the tail-suspended, hindlimb unloaded rat model was used to simulate microgravity-induced cardiovascular deconditioning for 28 days (SUS group); and during the simulation period, another group was subjected to daily 1-hour dorso-ventral (-G(x)) gravitation provided by restoring to normal standing posture (S + D group). The activity of vascular L-RAS was evaluated by examining the gene and protein expression of angiotensinogen (Ao) and angiotensin II receptor type 1 (AT1R) in the arterial wall tissue. The results showed that SUS induced an increase in the media thickness of the common carotid artery due to hypertrophy of the four SM layers and a decrease in the total cross-sectional area of the nine SM layers of the abdominal aorta without significant change in its media thickness. And for both arteries, the most prominent changes were in the innermost SM layers. Immunohistochemistry and in situ hybridization revealed that SUS induced an up- and down-regulation of Ao and AT1R expression in the vessel wall of common carotid artery and abdominal aorta, respectively, which was further confirmed by Western blot analysis and real time PCR analysis. Daily 1-hour restoring to normal standing posture over 28 days fully prevented these remodeling and L-RAS changes in the large elastic arteries that might occur due to SUS alone. In the ex-vivo experiments, to elucidate the important role of transmural pressure in vascular regional remodeling and differential regulation of L-RAS activity, we established an organ culture system in which rat common carotid artery, held at in-vivo length, can be perfused and pressurized at varied flow and pressure for 7 days. In arteries perfused at a flow rate of 7.9 mL/min and pressurized at 150 mmHg, but not at 0 or 80 mmHg, for 3 days led to an augmentation of c-fibronectin (c-FN) expression, which was also more markedly expressed in the innermost SM layers, and an increase in Ang II production detected in the perfusion fluid. However, the enhanced c-FN expression and increased Ang II production that might occur due to a sustained high perfusion pressure alone were fully prevented by daily restoration to 0 or 80 mmHg for a short duration. These findings from in-vivo and ex-vivo experiments have provided evidence supporting our hypothesis that redistribution of transmural pressures might be the primary factor that initiates region-specific remodeling of arteries during microgravity and the mechanism of IAG is associated with an intermittent restoration of the transmural pressures to their normal distribution. And they also provide support to the hypothesis that L-RAS plays an important role in vascular adaptation to microgravity and its prevention by the IAG countermeasure.

[Chronic blockade of angiotensin II type 1 receptor cannot completely prevent structural adaptation in vessels of simulated weightless rats].

Our previous studies suggest that the vascular local renin-angiotensin system (L-RAS) plays a pivotal role in the region-specific vascular adaptation due to simulated weightlessness. The present study was designed to determine whether simulated weightlessness still induced adaptive changes in rat vessels when angiotensin II type 1 receptor (AT(1)R) was chronically blocked by the administration of losartan, and whether the expressions of key elements in the L-RAS in the large arteries would change. Tail suspension for 4 weeks was used to simulate the physiological effect of weightlessness. The responses of the basilar, anterior tibial, carotid arteries and abdominal aorta were observed by morphometric technique with light microscopy. The expressions of angiotensinogen (AGT) and AT(1)R in the walls of common carotid artery and abdominal aorta were determined using immunohistochemical technique. The results showed that simulated weightlessness induced hypertrophy of the media of basilar artery and smooth muscle layers of carotid artery, but atrophic change in the anterior tibial artery and abdominal aorta. After 4 weeks of losartan treatment, all these arteries showed significant atrophic changes. However, simulated weightlessness still induced relative hypertrophy of the basilar artery and carotid artery and atrophy of the abdominal aorta when AT(1)R was blocked. After 4 weeks of simulated weightlessness, the expressions of AGT and AT(1)R were upregualted in the wall of carotid artery, but downregulated in the wall of abdominal aorta and perivascular tissues. Losartan decreased AGT and AT(1)R expressions only in the wall of abdominal aorta; whereas simulated weightlessness further decreased AT(1)R expression in the wall of abdominal aorta when AT(1)R was blocked. We conclude that simulated weightlessness for 4 weeks still induces structural changes and upregulates or downregulates the key elements in L-RAS in the large and medium-sized arteries from fore and hind body parts of rats when AT(1)R is blocked. The results suggest that the L-RAS in arterial tissue plays a pivotal role in these differential structural changes. However, there still exist other regulatory pathways to mediate the adaptive regulation of cerebral vessels when AT(1)R is blocked.

Development of a phage displayed disulfide-stabilized Fv fragment vaccine against Vibrio anguillarum.

Anti-idiotype monoclonal antibody 1E10 can mimic the protective epitope of Vibrio anguillarum and be used as vaccine to prevent fish infection of V. anguillarum. In this study, the variable heavy (V(H)) domain and variable light (V(L)) domain of mAb1E10 were cloned by RT-PCR and were linked to each other by a disulfide bond engineered at position 44 of V(H) and position 105 of V(L) that lie between structurally conserved framework positions. Mutated V(H) 44 and V(L) 105 were inserted into phagemid pCANTAB5E. When co-transfected by recombinant pCANTAB5E and helper phage M13KO7, the host Escherichia coli cells secreted disulfide-stabilized Fv fragment (dsFv) which displayed on the surface of filamentous phage. The binding specificity of the phage-displayed dsFv was proved by ELISA method. Protection experiment showed that Japanese flounders can develop high titer of antibody against the dsFv and survival ratio of vaccinated group was significantly different from control groups. Thus, this phage-displayed dsFv may be used as vaccine against V. anguillarum in fishery.

[Cloning and sequence analysis of variable region gene of anti-idiotype monoclonal antibody against Vibrio alginolyticus].

AIM: To clone and sequence V(H) and V(L) genes of anti-idiotype monoclonal antibody (mAb) against vibrio alginolyticus. METHODS: Total RNA was extracted from hybridoma cell AL1 secreting mAb against vibrio alginolyticus and cDNA was amplified by RT-PCR. Then the cDNA was inserted into PMD18-T vector and its sequence was analyzed. RESULTS: The V(H) gene contained 369 bp and encoded 123 amino acid residues; the V(L) gene contained 339 bp and encoded 113 amino acid residues. There were four FRs, three CDRs and two characteristic cysteine residues in the V(H) and V(L) genes, respectively. CONCLUSION: The successful cloning of the V(H) and V(L) genes of anti-idiotype mAb against vibrio alginolyticus provides a sound basis for construction of gene-engineering vaccine of the anti-idiotype mAb against vibrio alginolyticus.

Distribution, cloning and sequencing of GnRH, its receptor, and effects of gastric acid secretion of GnRH analogue in gastric parietal cells of rats.

Our objective was to study the distribution of gonadotropin-releasing hormone (GnRH) and its receptor, cloning and sequencing of GnRH and its receptor gene in cultured gastric parietal cells of rats. The distribution of GnRH and its receptor mRNA were investigated through immunocytochemical ABC methods and in situ hybridization methods in cultured gastric parietal cells of rats. After isolation of the total RNA from the parietal cells, RT-PCR was conducted to obtain GnRH and its receptor cDNA. Then, the products of PCR was purified, digested by the restriction enzyme of Hind III and EcoR I, and DNA fragments of interests were cloned into pUC19 vector. The products of PCR were analyzed by sequencing with Sanger's method after identified by PCR and digestion of restriction enzyme. Gastric parietal cells showed GnRH and its receptor immunoreactivity; positive material was located in cytoplasm other than in nuclei. GnRH and its receptor mRNA hybridized signals were also detected in cytoplasm with negative nuclei. The specific amplified band of GnRH and its receptor sequences were detected through Agarose gel electrophoresis, and GnRH gene sequence is identical to that of GnRH which has been reported in rat hypothalamus and GnRH receptor sequence is identical to that of the pituitary of rat. GnRH analogue (Alarelin) could inhibit the gastric acid secretion both by direct actions on parietal cells and by inhibiting vagous function. Our data suggest that GnRH could be produced by gastric parietal cells of rats and may modulate physiological function of gastric parietal cells of rats through autocrinal and paracrinal way.

Expression of gonadotropin-releasing hormone receptor and effect of gonadotropin-releasing hormone analogue on proliferation of cultured gastric smooth muscle cells of rats.

AIM: To investigate the expression of gonadotropin-releasing hormone (GnRH) receptor and the effects of GnRH analog (alarelin) on proliferation of cultured gastric smooth muscle cells (GSMC) of rats. METHODS: Immunohistochemical ABC methods and in situ hybridization methods were used to dectect protein and mRNA expression of GnRH receptor in GSMC, respectively. Techniques of cell culture, OD value of MTT test, measure of (3)H-TdR incorporation, average fluorescent values of proliferating cell nuclear antigen (PCNA) and flow cytometric DNA analysis were used in the experiment. RESULTS: The cultured GSMC of rats showed immunoreactivity for GnRH receptor; positive staining was located in cytoplasm. GnRH receptor mRNA hybridized signals were also detected in cytoplasm. When alarelin (10(-9), 10(-7), 10(-5) mol/L) was administered into the medium and incubated for 24 h, OD value of MTT, (3)H-TdR incorporation and average fluorescent values of PCNA all decreased significantly as compared with the control group (P<0.05). The maximum inhibitory effect on cell proliferation was achieved a concentration of 10(-5) mol/L and it acted in a dose-dependent manner. Flow cytometric DNA analysis revealed that alarelin could significantly enhance ratio of G(1) phase and decrease ratio of S phase of GSMC of rats (P<0.05). The maximum inhibitory effect on ratio of S phase was at the concentration of 10(-5) mol/L and also acted in a dose-dependent manner. CONCLUSION: Our data suggest that GnRH receptor can be expressed by GSMC of rats. GnRH analogue can directly inhibit proliferation and DNA synthesis of rat GSMC through GnRH receptors.

[Localization of the estrogen receptor alpha and beta-subtype in the nervous system, Hatschek's pit and gonads of amphioxus, Branchiostoma belcheri].

Immunocytochemical localization were investigated in the nervous system, wheel organ, Hatschek's pit and gonads of amphioxus using polyclonal antibodies against estrogen receptor-alpha and beta. The results revealed that ER-alpha and beta protein distributed in the above regions in larvae and adult at different developmental stages of both sexes. A major ER-alpha were expressed within nucleus of nerve cells, a few expressed in the cytoplasm and process as well as fiber of nerve cells in the forebrain, midbrain, hindbrain and nerve tube, while ER-beta were detected in the cytoplasm or on the cellular membrane, a few were within nucleus. ER-alpha immunopositive material distributed mainly in the nucleus of epithelial cells in the second layer of Hatschek's pit, a few were in the cytoplasm of upper layer epithelial cells, while ER-beta distributed in the nucleus of upper layer. In gonads, ER-alpha were distributed in the cytoplasm and nucleolus of oogonia and oocyte of small growth stage, germinal vesivle(nucleus) showed immunonegative reaction. In the large growth stage, strong immunopositive reaction were showed in nuclear membrane and nucleolus of oocyte, and within nucleus of mature egg cell in the mature stage. ER-beta immunopositive material distributed in the cytoplasm of oogonia and early oocytes as well as egg envelope of mature egg cell, the germinal vesicle showed immunonegative reaction. In testis, both ER subtype were localized in the cytoplasm of spermatogonia, primary and second spermatocyte as well as Sertoli cell, and within nucleus of spermatid cell, while spermatozoa showed immunonegative reaction. On the other hand, the results of double staining revealed that ER-alpha and beta numerously coexisted in a same cell, and fewly expressed in different cells in above regions. It is found for the first time that both estrogen receptor subtype, which mediated the regulate role of estrogen to neuroendocrine tissues in amphioxus, distributed extensively in amphiuoxus. The different localization of ER-alpha and beta receptor in the target cells suggests that the mediating estrogen signal line and the mechanism of gene transcription may be different.

Expression of LIM Protein KyoT in Adult Mouse Testis.

The KyoT expression in the adult mouse was reported here for further investigation on the functions of KyoT in adult mouse. To study the expression of mRNA and protein of KyoT, Northern blot, RT-PCR, Immunohistochemical SABC methods and in situ hybridization methods were used in the experiments. Two kinds of KyoT were expressed at high levels in testis of adult mouse, and KyoT immunore activity was mainly located in Leydig's cells. The reactive substance was distributed in cytoplasm rather than in nuclei. KyoT mRNA hybridization signals were also detected in cytoplasm of Leydig's cells rather than in nuclei. The spermatogenic cells and negative controls showed negative results. These results suggest that KyoT was expressed in testis of adult mouse and mainly located in Leydig's cells.