Objective To observe the expression of 5-HT receptor 14 subtype mRNAs in the rat retina. Methods RT-PCR technique was used. Specific oligonucleotide primers were synthesized based on the complementary DNA sequence for each of the 14 cloned rat 5-HT receptor subtypes. Expression of 5-HT 1A、5-HT 2A、5-HT 2C、5-HT 3 and 5-HT 7 receptor subtype mRNAs were detected. Results Positive band densities of various 5-HT receptor subtypes were different in retina. The most bands were formed by 5-HT 7 and 5-HT 2A receptor subtypes, the secondary prominent bands were formed by 5-HT 2C and 5-HT 3 receptor subtypes. PCR products from each positively-detected receptor subtype were subcloned into pCRⅡ vector for sequencing and found in correspond to published complementary DNA sequences. No positive bands existed of 5-HT 1B, 5-HT 1D, 5-HT 1E, 5-HT 1F, 5-HT 2B, 5-HT 4, 5-HT 5A, 5-HT 5Band 5-HT 6receptor subtypes. Conclusions There may exist varieties of 5-HT receptor in rat retina, including 5-HT 1A, 5-HT 2A, 5-HT 2C, 5-HT 3 and 5-HT 7 receptor subtypes. It means the retina serotoninergic system may be a common feature in mammals and, by the light of the 5-HT receptor subtypes, play an important role in mediating the transmission of rod sensitive signals.

Objective To observe the distribution of protein kinase C? subunit(PKC?) in the rat nervous system. Methods Immunocytochemical staining for PKC? by using specific antibody against PKC? was performed. Results Intensely and densely stained PKC? immunoreactive neurons were mainly observed in the cerebral cortex,hippocampus,amygdaloid complex,cerebellar cortex, ventral and dorsal cochlear nuclei, caudal spinal trigeminal nucleus and the superficial layers of the spinal dorsal horn.Conclusion\ PKC?\|containing neurons are widely located in the rat nervous system.These results provide morphological evidence for the functional roles of PKC? in the signaling transduction of the nervous system.\;[

Objective To study the in vivo expression patterns of caspase-3 in normally aged primate brain. Methods Fresh brain tissue was taken from 4-year-old and 20-year-old female rhesus monkeys,4 for each age group. In vivo distribution of caspase-3-positive cells and its protein levels in different brain regions were investigated by immunohistochemical and immunoblot means,respectively. Results In the frontal cortex,most neurons lacked detectable caspase-3 immunoreactivity,whereas low to moderate caspase-3 immunostaining was found mainly in pyramidal cells in CA1,CA3 and CA4 subdivisions of hippocampus. And in the cerebellum,a small number of Purkinje cells were strongly stained in their cytosol and dendrites,while their nuclei lacked staining. Age-related changes in caspase-3 expression pattern were not found. Immunostaining of motor cortex in aged monkeys localized strong caspase-3 immunoreactivity to a limited number of large pyramidal cells in layer V. Immunoblotting revealed the 32 000 caspase-3 progenitor in all three brain regions. There were no differences in caspase-3 expression levels as a function of either brain region or age of animals. Conclusions These results suggest that caspase-3 is constitutively expressed in matured primate brain and there is no significant age-related change in its expression.

Objective:Preparated from seeds of Brassica alba, Sinapine was determinated by RP-HPLC. Methods: Seeds of Brassica alba were shattered, defatted by ether, extracted by alcohol, and crystallized by potassium thiocyanate, then ethanol and water were used to recrystallize for several times. The determination was performed by reversed-phase high performance liquid chromatography(RP-HPLC), using ZORBAX SB-Aq column 4.6 mm?250 mm, 5 ?m. The mobile phase consisted of acetonitrile-0.08 mol/L KH2P04 (20:80) at a flow rate of 1.0 ml/min, and the detection wavelength was 326 nm. Results: Pale yellow needle-like sinapine crystal was obtained, and the yield rate was 0.82%. The purity of sinapine thiocyanate crystal was over 95%. Conclusion: Preparation sinapine by this solvent extraction method was simple, high-purity, high yield, and could be used for industrial production.

Objective To evaluate the effects and toxic side-effects of radiotherapy combined with xeloda or carboplatin respectively for locally advanced esophagus carcinoma.Methods 74 cases with locally advanced esophagus carcinoma were randomly allocated into two groups.One group received radiotherapy combined with Xeloda,500mg at a time,twice a day,taken orally from the beginning of radiotherapy till end of treatment.The second group received radiotherapy combined with Carboplatin by intravenous drip at 100mg a day,for 5 days,from the first week of radiotherapy.Results The overall response rate in the radiotherapy combined with Xeloda group was 91.4％ and 89.7％ in the radiotherapy combined with Carboplatin group.The difference between the two groups was not statistically significant(P ＞0.05 ).Kaplan-Meier survival analysis showed no statistically significant differences in survival between the two groups.The toxic side-effects of gastrointestinal tract,reaction myelosuppression and late radiation reaction were notably alleviated in the Xeloda group compared with the Carboplatin group.Conclusion Compared with Carboplatin,Xeloda combined with radiotherapy had less toxic side-effects for treatment of advanced esophagus carcinoma,with an equal local control rate,which worthy of clinical application.

Objective To observe the developmental changes of projection and termination of proprioceptive afferent fibers in the mouse spinal cord. Methods Parvalbumin (PV) immunohistochemistry was used to label the proprioceptive afferents. Single and dual immunofluorescence histochemistry were used to examine the growth pattern of proprioceptive afferents and their relationships with motoneurons in the spinal ventral horn (VH). The stained sections were observed under a confocal laserscanning microscope. Results PV-like immunoreactive (LI) proprioceptive fibers first appeared in the dorsal column on embryonic (E) day 14, then entered the gray matter on El5 and reached the intermediate gray matter and VH more obviously on E16. The number and intensity of PV-LI proprioceptive afferent fibers and punctata increased in the VH with age and reached a maximum during earlier postnatal (P) period (P0-P7). After P14, the number and intensity of proprioceptive afferents gradually decreased. The proprioceptive terminals seemed to form close relationship with motoneurons from E17. Conclusion The present study indicates that the somatotopic organization of proprioceptive afferents in the spinal cord is established during the late embryonic and early postnatal stages. These results provide evidence for understanding the development of the reflex movements.

The present study was designed to examine the developmental changes in projection and termination of nociceptive and proprioceptive afferent fibers in the spinal cord by labeling those two fibers with calcitonion gene-related peptide (CGRP) and parvalbumin (PV)separately in mouse embryos and neonatal pups aged embryonic day 15 to posanatal day 3 (E15 -P3). CGRP-like immunoreactive (LI)nociceptive fibers first appeared in the superficial dorsal horn (DH) at E16. The afferent projections extended laterally to the DH and entered into the deep portions of the DH at E17 and E18. After birth, the projection pattern of CGRP-LI fibers remained unchanged but the intensity of afferent terminals increased in the superficial laminae and their branching patterns became more complicated. In addition,CGRP-LI collaterals that projected into the contralateral DH were also examined after E16. Around birth, the contralateral projections were also found originated from the lateral part of the DH. PV-LI proprioceptive afferents were first observed entering the gray matter at E15 and reached the intermediate gray matter (IG) and the ventral horn (VH) more obviously on E16. The number and intensity of PV-LI fibers increased in the the VH with age and reached a maximum during earlier postnatal period ( P0-P3 ). The proprioceptive terminals seemed to form close relationship with motoneurons in the VH from E17. Our results indicate that the somatotopic organization of nociceptive and proprioceptive afferents in the spinal cord both are established during the late embryonic and early postnatal stages. These results help to understand the development of the sensory transmission in more details.

The present study was designed to examine the morphological pattern of primary afferent projections into the spinal dorsal horn by labeling the lumbar dorsal root ganglia with carbocyanine fluorescent dye DiI in mouse embryos and neonatal pups aged embryonic day 12 to postnatal day 3 (E12-P3). Primary afferent fibers projected into dorsal funiculus at E13, but did not penetrated into gray matter of dorsal horn until E15. The afferent projections became dense and entered the spinal gray matter more deeply at E16 and E17. By E18 the intensity of primary afferent in the deep part of the dorsal horn increased and their branching patterns became more complicated. Some of these primary fibers were also observed to ramify extensively in the superficial laminae. The projection pattern of primary afferent remained unchanged after birth, but the intensity of afferent terminals increased in the superficial laminae. In addition, afferent fiber collaterals that projected into the contralateral dorsal horn were also observed. They were first examined at E16 and mainly originated from the medial and deep part of the dorsal horn. Around birth, the contralateral projections were also found to originate from the lateral part of dorsal horn. Our results indicate that laminar organization of primary afferents in the spinal dorsal horn is established during the late embryonic and early postnatal stages.This organization then undergoes further refinement to match the pattern seen in the adult.

Immunocytochemical staining technique by using specific antibody against 5-HT1A receptor subtype (5-HT1AR) wasused to observe the distribution of 5-HT1AR immunoreactivity in the rat nervous system. The highest level of 5-HT1AR im-munoreactivity was observed in piriform cortex, septum, ventraldorsal thalamic nucleus, reticular thalamic nucleus, basolateralamygdaloid nucleus, Purkinje cell layer, red nucleus, facial nucleus and nucleus of the trapezoid body. Considerably weaker im-munoreactivity was detected in hippocampus, frontal cortex, mediodorsal thalamic nucleus, interpeduncular nucleus, mesen-cephalic trigeminal nucleus, dorsal raphe nucleus, spinal trigeminal nucleus, the superficial layers of the spinal dorsal horn, dor-sal root and trigeminal nerve ganglia, Very weak immunoreactivity was found in the olfactory bulb, caudate putamen,globus pal-lidus, nucleus diagonal band, bed nucleus stria terminalis, habenular nucleus, substantia nigra and superior olive. The presentresults indicate that 5-HT1AR immunoreactive structures are widely distributed in the rat nervous system and might play impor-tant role in mediating the multiple effects of 5-HT in the nervous system.

ObjectiveTo evaluate the clinical efficacy and safety of astragalus injection on the immune function in patients with senile sepsis.Methods Sixty patients with old age sepsis in Critical Care Medicine Department of Guangdong Provincial Traditional Chinese Medicine Hospital were enrolled and randomly assigned into control and treatment groups according to the table of random numbers, 30 cases in each group. According to 2012 sepsis guidelines for treatment, including antibacterial drug, mechanical ventilation, visceral function support, etc., the therapy was given to the control group; besides the treatment in the control group, intravenous drip of 60 mL astragalus injection(10 mL per ampoule) in 250 mL 0.9% normal saline was additionally given in the treatment group, once a day for 7 days. Before and after treatment, the immunological indexes, acute physiology and chronic health evaluationⅡ(APACHEⅡ) score, sequential organ failure assessment(SOFA) score, duration of mechanical ventilation and time of stay in intensive care unit(ICU), 28-day mortality and adverse drug reactions were compared between the two groups.Results Before treatment, there were no statistically significant differences in CD3+, CD3+CD4+, CD3+CD8+ and T helper cells /T suppressor cells(Th/Ts)levels between the two groups(allP>0.05), while CD3-NK+ of the control group was significantly higher than that in the treatment group〔(10.47±6.22)% vs. (6.26±4.13)%,P<0.05〕. After treatment in treatment group, CD3+, CD3+CD4+ and CD3-NK+ were increased, CD3+CD8+,Th/Ts were decreased compared with those before treatment; in the control group after treatment, CD3+,CD3+CD8+ and CD3-NK+ were decreased and CD3+CD4+ and Th/Ts increased compared with those before treatment. In the comparisons between the treatment group and control group after treatment, the differences in CD3+, CD3+CD4+ and CD3+CD8+ had statistical significance〔CD3+:(30.30±17.17)% vs.(41.91±22.29)%, CD3+CD4+:(31.54±13.24)% vs.(40.08±15.28)%, CD3+CD8+:(14.25±8.10)% vs.(9.52±9.33)%,allP<0.05〕; while the differences in Th/Ts and CD3-NK+ had no statistical significance(bothP>0.05). After treatment in the treatment group, IgG was increased compared with that in the control group〔IgG(g/L): 13.07±5.43 vs. 10.10±3.96,P<
0.05〕. The differences in IgA, IgM, complement(C3,C4) and total serum complement activity(CH50) in the comparisons between the two groups had no statistical significance after treatment(allP>0.05). The differences in APACHEⅡ score(13.83±6.18 vs. 15.90±7.48), SOFA score(7.38±4.66 vs. 6.89±4.19), time of stay in ICU(day: 11.63±5.13 vs. 13.62±8.08), invasive ventilation time(hour: 155.44±119.68 vs. 224.08±174.15) and noninvasive ventilation time(hour: 55.55±42.24 vs. 98.57±43.17) had no statistical significance in comparisons between the treatment group and control group after treatment(allP>0.05). The difference in 28-day mortality had no statistical significance in comparison between the treatment group and control group〔16.7%(5/30) vs. 20.0%(6/30),P>0.05〕. In 60 cases, there were 2 patients with adverse drug reaction, one diarrhea and another little rashes, the rest of the patients did not appear any drug side effect.ConclusionAstragalus injection combined with conventional western medicine therapy possibly has certain effect on adjustment of disturbance of immunologic functions in old patients with sepsis, and its therapeutic safety is well.