[Diversity and community characteristics of organic phosphate-mineralizing bacteria in the sea area between Minjiang Estuary to Pingtan.] / 闽江口-å¹³æ½­æµ·åŸŸæœ‰æœºè§£ç£·èŒå¤šæ ·æ€§åŠç¾¤è½ç‰¹å¾.

Microbial mineralization of organic phosphorus is an important component of marine phosphorus cycle. The research on organic phosphate-mineralizing bacteria (OPB) is helpful to reveal microbial driving mechanism of organic phosphorus mineralization in eutrophic sea area. The diversity and community characteristics of OPB were examined by Illumina high-throughput sequencing using the primer sets phoX in the sea area between Minjiang Estuary to Pingtan in April (spring) and July (summer) 2019. The results showed that the Shannon index of OPB in the surface seawater samples ranged from 3.21 to 7.91, and that the diversity at each station was greater in spring than that in summer. Shannon index of OPB in the sediment samples ranged from 2.04 to 8.70, which was greater in summer than that in spring. Shannon index of OPB in surface seawater of each station was higher than that of sediment in spring, while it was in adverse in summer. Nine phyla of OPB were detected in surface seawater, with Proteobacteria and Cyanobacteria being the most abundant. Tweleve phyla of OPB were detected in the sediments, with Proteobacteria and Bacteroidetes being the most dominant. OPB community composition at the genus level showed obvious spatio-temporal variation. Leisingera, Phaeobacter, Thalassococcus, and Pseudomonas were the major genera in the seawater in spring, while Synechococcus, Halioglobus, Roseovarius, Phaeo-bacter, Sulfitobacter, and Hyphomonas were the major genera detected in summer. Leisingera, Phaeobacter, Vibrio, and Sulfitobacter were major genera in the sediment in spring, while Azospirillum, Aminobacter, Sulfurifustis, Burkholderia, and Thiohalobacter were the major genera in summer. A large number of unclassified OPB were detected in both surface seawater and sediment. The redundancy analysis results showed that dissolved oxygen, water temperature, pH, dissolved inorganic nitrogen, NO2--N, and NO3--N had great influences on community distribution of OPB in the surface seawater. The abundant OPB in the surface seawater and sediment might play an important role in phosphorus cycle in this sea area.

Clinical and mechanism research on functional erectile dysfunction treated with moxibustion and / 中国针灸

OBJECTIVE@#To observe the clinical therapeutic effect on functional erectile dysfunction (FED) of kidney deficiency and liver stagnation treated by @*METHODS@#A total of 120 patients with FED were randomized into an observation group (60 cases, 2 cases dropped off) and a control group (60 cases, 4 cases dropped off). In the control group, the patients were treated with oral @*RESULTS@#After treatment, the scores of IIEF5, EHS and EDITS were all increased as compared with the values before treatment (@*CONCLUSION@#The combined therapy of

[Microstructural characteristics of lymphatic vessels in skin tissues of acupoints "Taichong" and "Yongquan" in the rat].

OBJECTIVE: To provide a new method for investigating the histological characteristics of acupoints by obser-ving the microstructure of the lymphatic vessels in the skin tissue of "Taichong" (LR3) and "Yongquan" (KI1) regions. METHODS: Six male SD rats were used in the present study. The skin tissue of LR3 and KI1 from the hind foot were taken following transcardial perfusion with 4% paraformaldehyde. The skin tissues were cut into sagittal sections with a freezing microtome and stained by fluorescent immunohistochemistry with lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), calcitonin gene-related peptide (CGRP), and phalloidin for displaying the lymphatic vessels, nerve fibers, and blood vessels, separately. The samples were viewed and recorded using fluorescent microscope and laser scanning confocal microscope. RESULTS: In the skin tissue of LR3 and KI1 regions, the lymphatic vessels, nerve fibers, and blood vessels were labeled with LYVE-1, CGRP and phalloidin, respectively. The lymphatic capillaries were found to start from the enlarged blind end and distribute in the dermis and subcutaneous tissues with various forms, crisscrossing. Abundant blood capillaries at various thickness distributed around the lymphatic capillaries in a parallel or crossed pattern, intermingled with free nerve fibers. CONCLUSION: The lymphatic capillaries, blood capillaries and nerve fibers extensively distribute in the skin tissues of LR3 and KI1 regions in rats, suggesting an involvement of the immunomodulation in the effects of acupuncture in pathological conditions, despite being not limited to the acupoint regions in the distribution of lymphatic capillaries.

[Review on application of neural tracing technique to experimental research of acupuncture].

OBJECTIVE: To collect literatures on the application of neural tracing technique in experimental acupuncture research, and summarize its application status in acupoints, meridians and Zang-fu organs. METHODS: We collected papers published from databases of CNKI (1979-2017), Wan-Fang (1990-2017), VIP (1989-2017) and PubMed(1997-2017)by using keywords of "neural tracing technique" "neuroanatomic tracing" "neural tracers" and "acupuncture" "electroacupuncture" "auricular acupuncture" "eye acupuncture" "meridians" "acupuncture points" "acupoint injection", and made a summary about the current state of application of neural tracing technique in the fields of acupoint, meridian and Zang-fu organs (viscera). RESULTS: A total of 94 articles were collected, the most commonly used neural tracers were horseradish peroxidase and cholera toxin subunit B. The experimental animals used were rat, rabbit, cat, monkey, etc., and injection site was acupoint. After the injection of neural tracers, the survival time of animals was range from 1 day to 12 weeks, and the labelled tissues included neurons and nerve fibers of the sensory, motor, and autonomic systems. The outcomes of neural labeling mainly revealed the segmental pattern, neuroanatomical connection (neural pathways/circuits) and chemical features (shown by immunohistochemical staining) of neurons and nerve fibers innervating both the acupoints and visceral organs, suggesting their involvement in the effect of acupuncture and moxibustion treatment. CONCLUSION: This application of neural tracing technology help us understand the under-lying mechanisms of acupuncture and moxibustion interventions from different perspectives of neural pathways/circuits and related chemical properties, which also lays a greater role for this technology in future experimental acupuncture research.

[Distribution of serotonin-positive cells in local cutaneous tissues of different acupuncture points at hindlimbs in rats].

OBJECTIVE: To investigate the expression of serotonin (5-HT) in local tissue of different acupoint regions at the hind limbs of rats. METHODS: A total of 6 male SD rats were used in the present study. The local tissues of "Neiting"(ST44), "Zusanli"(ST36) and "Futu"(ST32) at the hindlimbs were dissected after transcardiac perfusion with fixative solution containing 4% paraformaldehyde, phosphate buffer, etc., fixed again in 4% paraformaldehyde and cut into sections (at 20 µm thick). Then, the tissue sections were subjective into immunofluorescence stain for displaying 5-HT positive cells and calcitonin gene-related peptide (CGRP) -positive fibers, vessel-like structure (labelled with phalloidin) and cellular nucleus (labelled with DAPI), separately. After that, the tissue samples were observed and recorded using a laser confocal microscope. RESULTS: The 5-HT labeled cells presented flat or round form in the local tissues of the 3 acupoint regions. These cells were mainly located in the dermis and subcutaneous tissue, and concentrated around the vessel-like structure and CGRP-positive nerve fibers. The number of cutaneous 5-HT positive cells was most in ST44 (156.6±12.1), and fewer in ST36 (83.5±18.2)and ST32(53.3±5.2)in turn. CONCLUSION: 5-HT-positive cell is an important member in local tissue of acupoints (ST44, ST36 and ST32), and varies in number according to their locations, suggesting an involvement of different densities of 5-HT-positive cells in different levels of regulatory effects of acupuncture stimulation of these acupoints.

[Moxa-stick moxibustion activates mast cells of small intestine tissue in rats].

OBJECTIVE: To explore the effect of moxa-stick moxibustion and joss-stick moxibustion at "Guanyuan" (CV4) on the activity of mast cells in the small intestine tissue in rats. METHODS: Twelve male SD rats were randomly divided into control, joss-stick moxibustion and moxa-stick moxibustion groups (n=4 rats in each group). Joss-stick or moxa-stick moxibustion was applied to CV4 for 10 min. After moxibustion, the skin temperature of the CV4 region was measured immediately with a thermometer. The mast cells and nerve fibers in the small intestine tissue were displayed by immunofluorescence histochemistry. RESULTS: Compared with the control group, the skin temperature of the CV4 region in both the joss-stick and the moxa-stick groups were significantly increased (P<0.05), while the skin temperature of the moxa-stick group was significantly higher than that in the joss-stick group (P<0.05). There were a large number of tryptase-positive mast cells in the small intestine of rats, some of which were co-expressed with lysosomal-associated membrane protein 1, displaying an activated state. The average numbers of mast cells in the control, joss-stick and moxa-stick groups were 9.2±3.6, 10.8±5.3 and 17.1±6.3, respectively, being significantly higher in the moxa-stick group than in the control and joss-stick groups (P<0.05). In addition, calcitonin gene-related peptide(CGRP) and neuropeptide Y(NPY) positive nerve fibers were found around the mast cells in the small intestinal tissues. CONCLUSION: Moxa-stick moxibustion gives rise a higher temperature at CV4 to activate mast cells surrounded by CGRP and NPY positive nerve fibers in the small intestine tissue in rats, suggesting an involvement of the sensory and sympathetic nervous system in the activation of intestinal mast cells possibly by way of somatic sympathetic reflex.

[Community characteristics of polyphosphate accummulating organisms in the heart sediment of three reservoirs in Fujian Province, China.] / 福建省3座水库库心沉积物聚磷菌的群落特征.

Polyphosphate accummulating organisms (PAOs) play an important role in the phosphorus metabolic cycling in the sediment of reservoir. We assessed the diversity and community structure of PAOs in the sediments by T-RFLP and clone sequencing which targeted ppk1 gene at the hearts of three reservoirs (Jiulongjiangxipi reservoir, Sanshiliujiao lake reservoir and Dongyaxi reservoir) in Fujian Province. The results showed that the diversity of PAOs varied among different reservoirs, though not statistically significant. The diversity of PAOs in the Sanshiliujiao lake reservoir was highest (Shannon index H=2.89±0.03, Simpson index D=0.06±0.01). The community structure of the PAOs in the Sanshiliujiao lake reservoir was most complicated, consistent with the results of the T-RFLP. The differences of dominant PAOs genera in three reservoirs were distinct, mainly concentrated in the Proteobacteria, Actinobacteria and Acidobacteria. The percentage of those three phylum accounted for 74.5%, 85.0% and 75.0%, respectively, of the total PAOs. The dominant groups in each reservoir sediment were Anaeromyxobacter and Solibacter. Various forms of phosphorus had certain influence on the diversity of PAOs. There were significantly correlation between Fe/Al-P and PAOs diversity and community structure. The dominant genus in the three reservoirs, Anaeromyxobacter, was positively correlated with all forms of phosphorus and significantly correlated with insoluble phosphorus such as OP and Ca-P, while Solibacter was negatively correlated with all forms of phosphorus. The results suggested that PAOs had important impacts on the phosphorus cycle of sediment in eutrophicatied reservoirs.

[Neuronal Correlation Between Acupoint BL 23 and the Adrenal Gland in Rats].

OBJECTIVE: To explore the neuroanatomical basis of acupoint-visceral correlation by studying the distribution of the neurons associated with acupoint "Shenshu" (BL 23) area and adrenal gland in rats. METHODS: AF 488-CTB and AF 594-CTB were injected into the left side of BL 23 area and adrenal gland in the same rat respectively. Three days after injection, the dorsal root ganglions (DRG), sympathetic chain, and spinal cord were dissected out from the perfused rats. The neuronal labeling with AF 488/594-CTB was directly observed on the sections under a fluorescent microscope or a laser scanning confocal microscope. RESULTS: All neural labeling was observed in the injection side. The sensory neurons associated with both acupoint BL 23 and adrenal gland distributed from thoracic (T) 10 to lumbar (L) 2 DRG with high concentration in T 12-T 13 and T 11-T 12, respectively, in which some of them were simultaneously labeled with both AF 488/594-CTB and located in T 12-L 1 DRG. For the sympathetic innervation, the postganglionic neurons correlated with BL 23 and adrenal gland were labeled with AF 488/594-CTB separately in the sympathetic chain at the lumbar segments, while the labeled preganglionic neurons were only observed at the lateral horn of T 11-T 13 spinal segments in the cases of adrenal gland. In addition, the labeled motor neurons were mainly detected in the spinal ventral horn at cervical (C) 7-C 8 and T 11-L 1 segments. CONCLUSION: These results indicate that there are segmental correlation between BL 23 and adrenal gland on the sensory and sympathetic innervations, and this correlation might be a neural pathway for modulating the function role of adrenal gland through BL 23 needling.

Differential innervation of tissues located at traditional acupuncture points in the rat forehead and face.

OBJECTIVES: To compare the neural pathways associated with the tissues located at different traditional acupuncture points in the rat forehead and face using the cholera toxin B subunit (CTB) neural tracing technique. METHODS: After injection of CTB into the tissues at GB14, ST2 and ST6 in the rat, the neural labelling associated with each acupuncture point was revealed by fluorescent immunohistochemistry of the nervous system, including the trigeminal ganglion (TRG), cervical dorsal root ganglia (DRG), spinal cord and brain. RESULTS: The CTB labelling included sensory neurons and their transganglionic axonal terminals, as well as motor neurons. The labelled sensory neurons associated with GB14, ST2 and ST6 were distributed in both the TRG and cervical DRG, and their centrally projected axons terminated in an orderly fashion at their corresponding targets in the spinal trigeminal nucleus and cervical spinal dorsal horn. In addition, labelled motor neurons were observed in the facial motor nucleus, trigeminal motor nucleus and cervical spinal ventral horn, in which facial motor neurons projected to the tissues located at all three acupuncture points. Trigeminal motor neurons innervated both ST2 and ST6, while spinal motor neurons only correlated with ST6. CONCLUSIONS: These results indicate that the tissues located at each of these three traditional acupuncture points in the rat forehead and face has its own sensory and motor connection with the nervous system in a region-specific pattern through distinct neural pathways. Understanding the neuroanatomical characteristics of acupuncture points from the peripheral nervous system to the central nervous system should help inform acupuncture point selection according to the demands of the clinical situation.

[Our Considerations About Studies on Structure of Acupuncture Points].

The anatomical structure of acupuncture points is the carrier for needling stimulation induced neurobiological effects. Although researchers have never stopped investigating its structure and experienced different stages from macroscopic to microcosmic morphological material basis, our understanding still largely lags behind the requirements of the times. In the present paper, we review development of researches on acupuncture points in recent decades from 1) macroscopic anatomical structure including skin, bones, muscles, nerves, blood vessels, etc.; 2) macro-microcosmic structure as the thickness of skin and muscles, distribution of blood vessels and nerve fibers, and innervation; 3) microstructure including cell categories (mast cells, fibroblasts, macrophages, etc.), peptidergic nerve fibers, various receptors, chemical substances, and their relationship with the brain and immune-endocrine system (cutaneous-neuro-immune-endocrine-mental biological communication), etc. These research outcomes will not only help us demonstrate the acupoint's structure better at the level of the body surface and regional anatomy, but also, at the microstructural level, reveal different kinds of cells and their chemical compositions in its structure as well as their relationship with the regulatory system of the whole body. From the perspective of the cutaneous and systemic neuro-immuno-endocrine networks, it undoubtedly prompts us to profoundly explore the underlying biological mechanisms of acupuncture. To observe the regional mutual connection and interaction of different kinds of cells and their chemical mediators and molecules in the acupuncture point will improve our understanding about the nature of acupuncture points from the static to the dynamic point of view. This might be a new approach for us to further reveal the functional roles of acupuncture points.

Improving the Application of High Molecular Weight Biotinylated Dextran Amine for Thalamocortical Projection Tracing in the Rat.

High molecular weight biotinylated dextran amine (BDA) has been used as a highly sensitive neuroanatomical tracer for many decades. Since the quality of its labeling was affected by various factors, here, we provide a refined protocol for the application of high molecular weight BDA for studying optimal neural labeling in the central nervous system. After stereotactic injection of BDA into the ventral posteromedial nucleus (VPM) of the thalamus in the rat through a delicate glass pipette, BDA was stained with fluorescent streptavidin-Alexa (AF) 594 and counterstained with fluorescent Nissl stain AF500/525. On the background of green Nissl staining, the red BDA labeling, including neuronal cell bodies and axonal terminals, was more distinctly demonstrated in the somatosensory cortex. Furthermore, double fluorescent staining for BDA and the calcium-binding protein parvalbumin (PV) was carried out to observe the correlation of BDA labeling and PV-positive interneurons in the cortical target, providing the opportunity to study the local neural circuits and their chemical characteristics. Thus, this refined method is not only suitable for visualizing high quality neural labeling with the high molecular weight BDA through reciprocal neural pathways between the thalamus and cerebral cortex, but also will permit the simultaneous demonstration of other neural markers with fluorescent histochemistry or immunochemistry.

[Characteristics of Distribution of Blood Vessels and Nerve Fibers in the Skin Tissues of Acupoint "Taichong" (LR 3) in the Rat].

OBJECTIVE: To investigate the microstructure (blood vessels and nerve fibers) of the skin tissue in "Taichong" (LR 3) region for reveling morphological characteristics of acupoint. METHODS: Five SD rats were used in the present study. The skin tissue in the region of acupoint LR 3 was taken from the dorsum of hind foot following transcardial perfusion with 4% paraformaldehyde. Then, the skin samples were sagittally or horizontally cut into sections (20 µm or 40 µm in the thickness) to be stained with Phalloidin and calcitonin gene related peptide (CGRP) by using fluorescent histochemistry and immunohistochemistry. The labeled vascular structure and nerve fibers were observed and recorded using fluorescent microscope and laser scanning confocal microscope. RESULTS: In the skin tissue of LR 3, different types of blood vessels labeled by phalliodin, including capillaries and glomera mainly distributing in the superficial layer of the dermis, and thicker blood vessels and their sub-branches mainly existing in the deeper layer of the dermis and the subcutaneous layer, were found. In addition, CGRP positive nerve fibers were found to run parallel to the thin blood vessels or to gather around the thicker blood vessels. From the subcutaneous la-yer to the epidermis, blood vessels and nerve fibers coexisted and formed a stereo-network structure. CONCLUSIONS: In the skin of LR 3 area, there exists a stereo-network structure consisting of different types of blood vessels and nerve fibers. Although this structure is not specifically associated with acupoint area alone, this result may provide a new sight to further understand the microstructure of acupoint.