Serotonin (5-HT) neurons in the nucleus raphe magnus (NRM) of the rat were identified by light and electron microscope immunocytochemistry methods. Serotoninlike immunoreactive (5-HT-li) neurous were large multipolar and fusiform cells which mainly located in the ventrocaudal part of the NRM. With electron microscopic immunocytochemistry the following findings were revealed: 1. Immunostaining was present in the cytoplasmic matrix, outer membrane of mitochondria, rough endoplasmic reticulum, multivesicular bodies and vesicles. Some nuclei of the cells were stained. 2.5-HT-li somata, dendrites and axons were postsynaptic to numerous unlabeled terminals which contained clear round vesicles (20-30 nm) or mixed with small granular vesicles (40-70 nm), large granular vesicles (90-110 nm) and flat clear vesicles. 3. The 5-HT-li axons were unmyelinated fibers, and the 5-HT-li axon terminals were scarce in the NRM. 4.5-HT-li axon terminals and dendrites abutting on capillaries and 5-HT-li dendrite-glial direct appositions were found in the NRM.These results revealed the ultrastructural characteristics of 5-HT-li neuron in the NRM. The variety of unlabeled terminals making contact with 5-HT-li somata and dendrites suggests that several neuronal system with possibly different transmitters may regulate 5-HT raphe-spinal neurons which may play integrative role in the NRM. Tae relationship between 5-HT neuron and the significance of the local microcirculation and the relationship between 5-HT neurons and glia cells were also discussed in the present paper.

Substance P-like immunoreactive (SP-li) cell bodies and processes and their synaptic relations with other neural elements in the nucleus raphe magnus (NRM) of rats were studied by electron microscope. The results showed that SP-li neurons are fusiform, oval and multipolar cells which distribute evenly over the nucleus. Immunostaining was present in the cytoplasmic matrix and membranes of mitochondria, endoplasmic reticulum and vesicles. Both unmyelinated and myelinated SP-li axons are found in the NRM. SP-li dendrites are numerous in the NRM. SP-li dendrites and somata are contacted by a variety of round, flat and granular vesicle-containing terminals. A central SP-li dendrite receivie convergent inputs from unlabelled axonal boutons is frequently seen in the NRM. SP-li terminals are presynaptic to the unlabelled dendrites, also some SP-li terminals are apposed or presynaptic to SP-li dendrites. Axo-axonic (SP-li) synapses were found in the NRM. The unlabelled presynaptic boutons contain clear round vesicles or mixed with granular vesicles. A central SP-li terminalis contacted by several unlabelled terminals is also found in the NRM.

BACKGROUND: Osteoporosis is a serious threat to the health and quality of life in the elderly. It is important to establish an ideal experimental animal model to study the etiology and treatment of osteoporosis.OBJECTIVE: To establish a rat model of osteoporosis induced by different dosages of retinoic acid, thus selecting the optimal dosage.METHODS: Eighty female Sprague-Dawley rats were randomly divided into control, low-, middle- and high-dosage groups based on body mass (n=20 per group), The rats in the latter three groups were induced with 80, 100, and 120 mg/(kg?d) retinoic acid via gastric lavage for 14 days.RESULTS AND CONCLUSION: Compared with the control group, the bone mineral density, number of osteoblasts and osteoclasts, and bone microarchitecture in the low-dosage group showed no significant changes, while there were significant decrease in the serum level of calcium and bone mineral density of femur, significant increase in the number of osteoclasts at the femur and significant changes in the femoral microarchitecture in the middle- and high-dosage groups, especially in the middle-dose group. To conclude, 120 mg/(kg?d) retinoic acid via gastric lavage for 14 days can induce a stable osteoporosis model in rats.

A total of 30 cats were used in this experiment.The nucleus raphe magnus wasinjected with 0.1～0.3?l of 25～50% HRP solution(RZ:2.4～3.1).Two or three daysafter operation,the animals were killed and then perfused with the fixing fluid.Thebrainstem and spinal cord were sectioned serially on a freezing microtome and treatedaccording to the method of retrograde axonal transport of horseradish peroxidase.Parts of the sections were counterstained with toloidine-blue.The labelled neuronswere examined microscopically under bright-field and dark-field illumination Thelabelled neurons were distributed as follows.After injections of nucleus raphe magnus,a few labelled neurons were foundin the medial part of parafascicular nucleus of the thalamus.A greater numbers oflabelled cells were consistently found in the ventrolateral area of the central greymatter of the midbrain.The dorsal nucleus of the midbrain raphe had a few labelled neurons.The HRPpositive neurons were found in the nucleus Darkschewitsch,mesencephalic reticularformation,nucleus reticularis gigantocellularis,nucleus reticularis paramedianus,nucleus vestibularis medialis and nucleus vestibularis lateralis.In some cases a fewlabelled cells were also seen in the posterior area of hypothalamus,nucleus posteriorcommissure,nucleus tegmentalis dorsalis,superior collicus,nucleus annularis,reticularformation of the pons,nucleus nervi facialis and nucleus gracilis.In summary,the present studies show that the afferent connections of nucleusraphe magnus are widely concerned with the structures of the pain control.Itsuggests that the nucleus raphe magnus is an important relay station for pain control.

Objective To observe the impacts of acupuncture and moxibustion on kelch like epichlorohydrin related protein 1(Keap1)and nuclear factor E2 related factor 2(Nrf2)signal pathway in kidney tissue of cisplatin(DDP)-induced kidney injury model mice,and to explain the protective mechanism of acupuncture and moxibustion on improving kidney injury caused by DDP.Methods Forty SPF male KM rats were randomly divided into 4 groups,with 10 rats in each group.One day before the start of treatment,the three groups of mice outside the blank group were intraperitoneally injected with cisplatin 10 mg·kg-1 according to their body weight,and the blank group was injected with the same dose of 0.9%NaCl solution.The model was established 24 hours later.Both acupuncture group and moxibustion group selected"Dazhui"(GV14),"Ganshu"(BL18),"Shenshu"(BL23)and"Zusanli(Housanli)"(ST36)for acupuncture and moxibustion respectively,once a day for 5 days.The other two groups were fixed every day without treatment.After fasting for 1 day,the contents of BUN,Scr,CysC and NGAL in serum and Keap1 and Nrf2 in renal tissue were detected by ELISA;Western blot and Real-time PCR were used to detect the protein expression and gene transcription of Keap1 and Nrf2 in the kidney tissue of mice in each group.Results Compared with the blank group,the content of Keap1 protein,protein expression and relative expression of mRNA in the model group increased(P<0.05),the content of Nrf2 protein,protein expression decreased(P<0.05),Nrf2 relative expression of mRNA increased(P<0.05);Compared with the model group,the content of Keap1 protein,the expression of Keap1 protein and the relative expression of Keap1 mRNA in the kidney of mice in the acupuncture and moxibustion groups decreased(P<0.05);Nrf2 protein content,protein expression and relative mRNA expression increased(P<0.05).Conclusion Acupuncture and moxibustion can improve the renal function of DDP renal injury model mice and enhance their antioxidant stress ability,so as to improve the renal injury caused by DDP chemotherapy.Its mechanism may be related to Keap1/Nrf2 signal pathway.

OBJECTIVE: Acute lung injury (ALI) is a serious respiratory dysfunction caused by pathogen or physical invasion. The strong induced inflammation often causes death. Tanshinone IIA (Tan-IIA) is the major constituent of Salvia miltiorrhiza Bunge and has been shown to display anti-inflammatory effects. The aim of the current study was to investigate the effects of Tan-IIA on ALI. METHODS: A murine model of lipopolysaccharide (LPS)-induced ALI was used. The lungs and serum samples of mice were extracted at 3 days after treatment. ALI-induced inflammatory damages were confirmed from cytokine detections and histomorphology observations. Effects of Tan-IIA were investigated using in vivo and in vitro ALI models. Tan-IIA mechanisms were investigated by performing Western blot and flow cytometry experiments. A wound-healing assay was performed to confirm the Tan-IIA function. RESULTS: The cytokine storm induced by LPS treatment was detected at 3 days after LPS treatment, and alveolar epithelial damage and lymphocyte aggregation were observed. Tan-IIA treatment attenuated the LPS-induced inflammation and reduced the levels of inflammatory cytokines released not only by inhibiting neutrophils, but also by macrophage. Moreover, we found that macrophage activation and polarization after LPS treatment were abrogated after applying the Tan-IIA treatment. An in vitro assay also confirmed that including the Tan-IIA supplement increased the relative amount of the M2 subtype and decreased that of M1. Rebalanced macrophages and Tan-IIA inhibited activations of the nuclear factor-κB and hypoxia-inducible factor pathways. Including Tan-IIA and macrophages also improved alveolar epithelial repair by regulating macrophage polarization. CONCLUSION This study found that while an LPS-induced cytokine storm exacerbated ALI, including Tan-IIA could prevent ALI-induced inflammation and improve the alveolar epithelial repair, and do so by regulating macrophage polarization.
Abietanes ; Acute Lung Injury/drug therapy* ; Animals ; Cytokine Release Syndrome ; Cytokines ; Inflammation/drug therapy* ; Lipopolysaccharides/toxicity* ; Macrophage Activation ; Macrophages ; Mice ; Triacetoneamine-N-Oxyl/pharmacology*