Experimental technicians are indispensable team to fulfill experimental teaching.With the process of medical experimental teaching reform and the foundation of functional experiment course,experimental technicians face more challenges.In the new experimental teaching mode,experimental technicians should modulate themselves from multiple aspects,in order to be more competent for the job.

Objective To observe the distribution of GABA-containing neurons revealed by GFP expression and the colocalization of the GFP with parvalbumin(PV) in the caudal spinal trigeminal nucleus(Vc),the glutamate decarboxylase 67-green fluorescence protein(GAD67-GFP) knock-in mice were used in the present study. Methods Double-labeled techniques were used by in situ hybridization combined with immunohistochemistry for GFP and double immunofluorescence histochemistry for GFP and NeuN(neuronal nuclei protein,a neuronal marker) or PV.The stained sections were observed under light microscope and a confocal laser-scanning microscope. Results 1.Over 90% of GFP-positive neuronal cell bodies in the laminae Ⅰ and Ⅱ of the Vc showed hybridization signals for GAD67 mRNA,and that almost all neuronal cell bodies with GAD67 mRNA signals were GFP-positive;2.GFP-positive neurons were mainly distributed in the laminae Ⅰ and Ⅱ of the Vc and the vast majority of them were small neurons.The considerable number of GFP-positive processes and somata were most densely observed in the lamina Ⅱ of the Vc.The proportion of GFP-positive neurons in the NeuN-labeled neurons of the Vc was about 19.4% and 24.3% in laminae Ⅰ and Ⅱ,respectively;3.Double-labeled neurons for GFP/PV are mainly found in laminae Ⅰ and Ⅱ of the Vc.The proportion of GFP/PV double-labeled neurons was about 62.4% and 12.8% of total population of PV-and GFP-positive neurons in laminae Ⅰ-Ⅱ of the Vc,respectively.Conclusion GABAergic neurons are mainly distributed in the laminae Ⅰ and Ⅱ of the Vc which related closely to transmission of the nociceptive primary afferent information,and the majority of PV-positive neurons are GABAergic neurons.

Objective The co existence of two kinds of neurotransmitters in the cell bodies or axon terminals and the relationship between a neurotransmitter and its relevant receptor were observed at the ultrastructural level. Methods The pre embedding double labeled immuno electron microscopic techniques were used by peroxidase immunohistochemistry combined with immunogold silver labeled method. Results In the sections of the striatum,a large number of the SP like immunoreactive(\|LI) axon terminals(immunoperoxidase reaction products)and SP receptor(SPR)\|LI neuronal cell bodies and dendrites(immunoglold silver grains)were seen in the present study.Some SP\|LI axon terminals were observed to form symmetric axon somatic or axon dendritic synaptic contacts with SPR\|LI neuronal cell bodies and dendritic profiles.On the other hand,in the sections of caudal spinal trigeminal nucleus,a substantial number of axon terminals of the two kinds of the vesicular glutamate transporters,the differentiation associated Na + dependent inorganic phosphate cotransporter(DNPI)and vesicular glutamate transporter of type1(VGluT1),were labeled by peroxidase immunohistochemistry and immunogold silver grains.Some axon terminals were labeled by both DNPI and VGLuT1 immunoreaction formed asymmetric synaptic contact with non labeled dendrite profile.Conclusion\ The pre embedding double labeled immuno electron microscopic technique has the advantage of higher sensitivity and greater preservation of the antigen in the many tissues.Thus,the present study favors the application of this technique to study the co existence of two kinds of the neurotransmitters in the cell bodies or axon terminals and analyse the relationship between a neurotransmitter and its relevant receptor in the field of the neuroanatomy.

Objective:To summary the antibacterial agent therapy adjustment performed by clinical pharmacists in the collabora-tive rounds. Methods:Combined with physicians, clinical pharmacists participated in the ward round and antibacterial agent selection for one case of hospital-acquired pneumonia, meanwhile, pharmaceutical care was performed for the patient. Results:Clinical pharma-cists chose appropriate antibacterial agents for the patient to cover the suspected pathogens and control the infection in time through par-ticipating in the ward round and paying attention to the disease course progress, medical history and medication history. Conclusion:Clinical pharmacists can enhance the quality of drug therapy through participating in the drug therapy, providing the proposal for the antibacterial agent use and performing pharmaceutical service for physicians and patients.

The possible relationship of vesicular glutamate transporters ( VGluT1 and VGluT2 ) - and 5-HT-like immunoreactive (LI) terminals with mesencephalic trigeminal nucleus (Vme) neurons in the rat was examined by using triple-immunofluorescence histochemistry and double-labeled electron microscopic immunohistochemistry. Under confocal laser-scanning microscope, many neuronal cell bodies of Vme showed phosphate-activated glutaminase (PAG) -LI and the vast majority of them were large pseudounipolar neurons. A considerable number of VGluT1 -LI and VGluT2-LI terminals were widely distributed in Vme, the density of VGluT2-LI terminals was higher than that of VGluT1-LI. 5-HT-LI axonal varicosities had dense distribution in Vme, and some 5-HT-LI terminals also showed immunoreactivity for VGluT2. Some VGluT1-LI, VGluT2-LI, 5-HT-LI and VGluT2/5-HT-LI terminals were frequently observed in close apposition to the cell bodies of Vme neurons showing PAG-LI. Under electron microscope, VGluT1/VGluT2-LI and 5-HT-LI were visualized with silver grains and peroxidase products, respectively. Some terminals in Vme showed both VGluT2- and 5-HT-LI, these dual labeled varicosities usually made asymmetric contact with Vme neurons. Synaptic terminals that showed VGluT1-LI was also observed, but no coexpression of VGluT1 and 5-HT was found in Vme. The present results suggest that in the transmission of the proprioceptive sensory information from the orofacial regions to the higher center, glutamate and 5-HT may play important roles on the regulation of Vme neurons through complicated integration.

Objective To investigate the distribution of the vesicular glutamate transporters (VGluTs), VGIuT1 and VGIuT2, in the spinal cord in multiple species, including rat, cat and monkey. Methods An immunohistochemical technique was used in the present study. Results VGluTs were observed in the gray matter of the spinal cord of rat, cat and monkey. VGluTl-immunoreactivity (VGluT1-IR) was most intense in the inner part of lamina Ⅱ(Ⅱi) and the medial parts of laminae Ⅲ-Ⅵ of the spinal dorsal horn; weak in lamina Ⅰ and outer part of lamina Ⅱ(Ⅱo); moderate in other parts in the rat and cat. The laminar distribution of VGluT1-IR in the spinal cord in the monkey is different from those in the rat and cat. The VGluT1-immunopositive product was intense in laminae Ⅰ, Ⅱi, Ⅲ and the medial parts of laminae Ⅳ-Ⅵ of the spinal dorsal horn; weak to absent in lamina Ⅱo and the lateral parts of laminae Ⅳ-Ⅵ of the dorsal horn. In the spinal anterior horn of monkey, the VGluT1-immunoreactive profiles were observed sparsely just in lamina Ⅸ, not in other laminae. At the same time, some scattered immunoreactive profiles were present in the white matter, particularly at the medial parts of the dorsal column in monkey, but not in rat and cat. On the other hand, VGluT2 immunoreactivity (VGluT2-IR) was moderately present throughout the gray matter, it was most dense in laminae Ⅰ-Ⅱ and Ⅹ, and weak in the medial parts of the deep spinal dorsal horn (laminae Ⅲ-Ⅵ) in the three species. The VGluT2-immunopositive product was very sparse in the lateral parts of the deep spinal dorsal horn of the monkey. In the lateral spinal nuclei of the rat, moderate VGluT2-immunopositive product could be seen, but not in the cat and monkey. In three species, both VGluT1- and VGluT2-IR showed a punctuate distribution. Conclusion The present results indicated that VGluT1 and VGluT2 were expressed in the spinal cord in all species examined but differentially distributed in a species-specific manner, and they might play an important role in the activities of the excitatory neurons in the spinal cord of rat, cat and monkey.

Previous studies showed that the Vodm-LRF-including the dorsomedial part of the subnucleus oralis of the spinal trigeminal nucleus and its adjacent lateral reticular formation--contained the second-order neurons on the central pathway of the trigeminal proprioceptive sensation of the rat and the "zone-shaped area"-including the caudolateral part of the supratrigeminal nucleus (Vsup-CL). The dorsomedial part of principal sensory trigeminal nucleus (Vpdm) and two newly found nuclei: the areaventral to the motor trigeminal nucleus (AVM) and the area dorsal to the superior olivary nucleus (ADO)-contained the third order neurons of this pathway. Parvalbumin (PV) is one of the calcium-binding proteins, In this pathway, many PV-like immunoreactive (PV-LI) neurons were observed in Vodm LRF and the "zone-shaped arena", hut there has been no reports so far regarding whether these PV-LI neurons are projection neurons responsible for the transmission of proprioceptive information or the interneurons serving the modulatory function, in the present study, our aim was to solve the problem by a double labeling study by using retrograde tracing method combined with immunofluorescence histochemistry. The results showed that: (1) following the unilateral Fluoro-Gold (FG) injections into the ventral posteromedial nucleus (VPM) of the thalamus and the separated parts of the "zone-shaped area", viz, Vpdm, ADO and AVM, many FG-labeled neurons were always found contralaterally in the "zone-shaped area" and ipsilaterally in the Vodm-LRF, respectively; (2) in either the "zone-shaped area" or the Vodm-LRF, a substantial number of the FG retrogradely labeled neurons showed PV-LI. In the Vsup-CL, Vpdm, AVM and ADO, about 57%, 55%, 11% and 4% of the neurons projecting to the VPM of the thalamus showed pV-LI, respectively. Of the total population of PV-LI neurons in the Vsup-CL, Vpdm. AVM and ADO, about 23%, 79%, 53% and 16% were labeled by FG, respectively. Most of these PV/FG double-labeled neurons were medium- to small-sized, round, piriform or irregular in shape. On the other hand, in the Vodm-LRF, approximately 33%, 34% and 50% of the neurons which projected to the Vpdm, AVM and ADO expressed PV-LI, respectively. The percentage of PV/FG double-labeled neurons to the total number of the PV-LI neurons in Vpdm, AVM or ADO were 26%, 17% or 11%, respectively, The present results indicated that PV might play an important role in the transmission of the trigeminal proprioceptive information of the rat from Vndm-LRF to the contralateral VPM of the thalamus through the "zone-shaped area".

Objective To set up a method for determining the content of Salvianolic acid B in Qianggan Syrup.Methods HPLC was adopted.A mixture of methanol-acetonitrile-1.5% formic acid(30:10:60) served as the mobile phase with a flow rate of 1.00 mL/min.The detection wavelength was 286 nm.Results The linear range was 0.145～1.747 ?g,r=0.999 9.The average recovery rate was 99.79%,RSD= 0.23%.Conclusion The method is highly effective,quick and sensitive,and suitable for quality control of the preparation.

Objective To observe the co-localization of phosphate activated glutaminase(PAG) and parvalbumin(PV) in neurons,and the connections between preprodynorphin-like immunoreactive(PPD-LI) or preproenkephalin(PPE)-LI terminals and coexistent neurons of PAG-LI and PV-LI in the mesencephalic trigeminal nucleus(Vme) of the rat. Methods In the present study,the immunofluorescence histochemical double-staining and triple-staining technique were used.The staining results were observed under a confocal laser scanning microscope. Results Many neuronal cell bodies through the whole rostrocaudal extent of Vme showed PAG-LI and PV-LI.The majority of them were large pseudounipolar neurons.A considerable number of PV-LI neurons expressed PAG-LI,cell counts showed that more than 95% of PV-I neurons were immunostained with PAG-LI.Confocal laser scanning microscope further revealed that axonal varicosities with PPD-LI or PPE-LI were found to make close contact with neuronal cell bodies of Vme showed both PAG-LI and PV-LI. Conclusion The present results indicated that the glutamate and PV are involved in the transmission of the trigeminal proprioceptive sensory singals from the Vme to the higher centers,simultaneously,dynorphin-and enkephalin-immuoreactive terminals may also exert modulatory influence on the primary afferent cell bodies of the Vme.;

Objective To investigate whether the serotonin(5\|HT)\|like immunoreactive axons synapse upon the second\|order neurons on the central pathway of the trigeminal proprioception in the rat. Methods Electron microscopic double\|labeling of retrograde tract\|tracing technique combined with immunohistochemistry. Results Following the injection of wheat germ agglutinin\|horseradish peroxidase (WGA\|HRP) into the dorsomedial part of the principal sensory trigeminal nucleus(Vpdm) of the rat and the immunostaining for 5\|HT, there were neurons retrogradely labeled by WGA\|HRP and axon terminals with 5\|HT\|like immunoreactivity(5\|HT\|LI) in the dorsomedial part of the subnucleus oralis of the spinal trigeminal nucleus and its adjacent lateral reticular formation(Vodm\|LRF).At the electron microscopic level,the 5\|HT\|immunostained axonal profiles were observed to make synaptic contacts with the WGA\|HRP\|labeled neurons.Both the axon\|somatic and axon\|dendritic synapses were found.These synapses were mainly of symmetric type although the asymmetric ones were also seen.Conclusion\ The present results provided a morphologic basis for the serotoninergic terminals which might be involved in the regulatory effect for the trigeminal proprioceptive information transmission.\;[