Responsive regularity of neurons in the hindlimb region of primary somatosensory cortex to different temperature thermal needle stimulation of "Zusanli"(ST36) in mice. / å°é¼ åˆçº§ä½“æ„Ÿçš®å±‚åŽè‚¢åŒºç¥žç»å ƒå¯¹"足三里"不同温度热刺激的响应规律.

OBJECTIVES: To study the regularity of central response to thermal needle stimulation of "Zusanli" (ST36) at different temperature, and to analyze the temperature difference of central responses. METHODS: Six male C57BL/6j adult mice were used in the present study. For observing activities of neurons in the hindlimb region of left primary somatosensory cortex (S1HL, A/P=0.46 mm, M/L=1.32 mm, D/V=-0.14 mm) by using a fast high-resolution miniature two-photon microscopy (FHIRM-TPM), the mice were anesthetized with 3% isoflurane (inhalation), with its head fixed in a stereotaxic apparatus, then, adeno-associated virus (AAV-hSyn-GCaMP6f-WPRE-hGHpA, for showing intracellular calcium transients in neurons transfected) was injected into the left S1HL region using a micro-syringe after scalp surgical operation. The mice's right ST36 were stimulated using internal thermal needles with the temperature being 43 ℃, or 45 ℃, or 47 ℃, separately. Image J software and MATLAB 2020b software were used to process the image data of neuronal calcium activity (Ca2+ signaling) in the left S1HL region, including the instant maximum calcium peak value (ΔF/F) in 2 s, instant calcium spike frequency in 2 s, short-term calcium peak value (ΔF/F) in 3.5 min, short-term calcium spike frequency in 3.5 min, calcium peak duration in 3.5 min, maximum calcium peak value (ΔF/F) at the 1st , 2nd and 3rd min, and calcium spike frequency at the 1st, 2nd and 3rd min after thermal needle stimulation. RESULTS: In comparison with the normal temperature needle stimulation, the instant intracellular maximum calcium peak value, instant calcium spike frequency, short-term maximum calcium peak value, short-term calcium spike frequency, and calcium peak duration of S1HL neurons in response to 43 ℃, 45 ℃ and 47 ℃ internal thermal needle stimulation of ST36 were significantly increased (P<0.001, P<0.01). Comparison among the 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation showed that the 45 ℃ thermal needle stimulation was obviously superior to 43 ℃ and 47 ℃ thermal needle stimulation in increasing instant calcium spike frequency, short-term calcium spike frequency and calcium peak duration of S1HL neurons (P<0.001, P<0.01). The 47 ℃ thermal needle stimulation was stronger than 43 ℃ and 45 ℃ thermal needle stimulation in increasing the instant maximum calcium peak value (P<0.001). The maximum calcium peak value was apparently higher (P<0.001) at the 2nd min than that at the 1st and 3rd min after 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation. No significant differences were found in the short-term maximum calcium peak value among the 3 thermal needle stimulation and in the calcium spike frequency among the 3 time points after 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation. CONCLUSIONS: S1HL neurons respond to all 43 ℃, 45 ℃ and 47 ℃ thermal needle stimulation of ST36 in mice, while more actively to 45 ℃ thermal needle stimulation.

Dynamics of RNA localization to nuclear speckles are connected to splicing efficiency.

Nuclear speckles, a type of membraneless nuclear organelle in higher eukaryotic cells, play a vital role in gene expression regulation. Using the reverse transcription-based RNA-binding protein binding sites sequencing (ARTR-seq) method, we study human transcripts associated with nuclear speckles. We identify three gene groups whose transcripts demonstrate different speckle localization properties and dynamics: stably enriched in nuclear speckles, transiently enriched in speckles at the pre-mRNA stage, and not enriched in speckles. Specifically, we find that stably-enriched transcripts contain inefficiently spliced introns. We show that nuclear speckles specifically facilitate splicing of speckle-enriched transcripts. We further reveal RNA sequence features contributing to transcript speckle localization, underscoring a tight interplay between genome organization, RNA cis-elements, and transcript speckle enrichment, and connecting transcript speckle localization with splicing efficiency. Finally, we show that speckles can act as hubs for the regulated retention of introns during cellular stress. Collectively, our data highlight a role of nuclear speckles in both co- and post-transcriptional splicing regulation.

[Analysis on developmental characteristics of acupoint theory].

The theory of acupoints has gone through a long process from its embryonic form to its maturity, including its gradual improvement in later generations. Starting from the acupuncture literature in different historical periods, we, in the present paper, gave some examples and analyzed the connotation of important concepts in the development of acupoint theory. It is believed that the signs of the establishment of acupoint theory mainly include the standardization of acupoints' names, the specific description of acupoint positioning, the enrichment and explicitation of the content of acupoints' indications, the highlights of the meridian theory via classification of acupoints, and the interpretation of connotations of acupoints from a clinical perspective.

[Parameters and connotations of quantity of mild moxibustion stimulation].

The quantity of mild moxibustion stimulation is an important factor affecting clinical therapeutic effect. In the present study, we collected related literature by using key word "moxibustion stimulation quantity" from CNKI Database and make a summary about its concept. The quantity of mild moxibustion stimulation has two common characters, namely thermal intensity and cumulative stimulating quantity, and contains 6 components (parameters), including thermal energy, thermal stimulus, heated area of the skin, and stimulating duration, intensity and frequency. According to the facts mentioned above, we hold that the mild moxibustion quantity actually has 3 dimensions, i．e., the duration of stimulation, the stimulating quantity of one treatment session, and the total stimulating quantity of multiple treatment sessions. Accurately grasping and rationally using the basic term of the quantity of mild moxibustion is not only conductive to control clinical moxibustion operation, but also conforms to the basic requirements for quantitative control of intervention measures in experimental researches.

[Could Huatuo Jiaji (EX-B 2) Acupoint Be Assigned to the Governor Vessel ?]

The meridian tropism of acupoints is an important academic way to recognize and deal with the relationship between acupoints and meridians in acupuncture medicine. The Huatuo Jiaji (EX-B 2) (Jiaji for short) acupoint family consisting of 34 points has long been considered to be an important extra-acupoint since the ancient times of China and is widely employed in clinical practice through continuous development in Chinese past generations. The present paper introduces 1) historical origin and development of EX-B 2, and 2) intercorrelation among the EX-B 2, spine and the Governor Vessel in anatomical positions. On the basis of these, we also discuss the possibility of that EX-B 2 is assigned to the Governor Vessel, its academic significance including common features and actions in physiological and pathological aspects and clinical significance in diagnosis and treatment of disorders.

[Discussion on Classical Morphological View of Acupoints: Taking Book A-B Classic of Acupuncture and Moxibustion as An Example].

After analyzing the contents of Volume Ⅲ of A-B Classic of Acupuncture and Moxibustion, we propose a concept of classical morphological view about acupoints in terms of their superficial signs and structure in Chinese ancient times. That is to say that the ancient medical experts' understanding about acupoints contain locality, layers and variability. These 3 aspects were applied to the classification and location of acupoints, as well as to the clinical treatment in the ancient times. Furthermore, the classical view about acupoints also contains the ideas of Shou-Xing (, keeping paying attention to needling the local acupoints only in the light of the focus) and Shou-Shen (, keeping paying attention to both the related acupoints and appropriate manipulations in the light of pattern identification of symptoms and signs as well as the patients' physical constitutions) in performing acupuncture treatment. Owing to the influence of the idea of Shou-Shen, the classical view about the signs and structure of acupoints presented a weakening trend gradually in ancient Chinese medical circle.

[Discussion on the cognition and development process of experimental animal acupoints].

To review the development history of the experimental animal acupoints. To analyze different historical periods, the names of animal acupoints, positioningmethods and references, it is believed that the understanding of experimental animal acupoints has gone through four stages, including initiation, accumulation, specification and standards. At the same time, some important events in the development of experimental animal acupoints were sorted out, including the publication of key papers, books, teaching materials, and the convening of important conferences. In addition, the significance of the above stages of development and the relationship between them were also analyzed to provide basic clues and evidence for the rational understanding of the characteristics and laws of experimental animals. It is concluded that the development of experimental animal acupoints has evolved from simple to complex, from fuzzy to precise.

[From on Banning of Acupuncture in Plain Questions to Acupuncture Prohibition：the formation of early acupuncture safety regulation].

Acupuncture safety is an unavoidable problem in acupuncture clinical. On Banning of Acupuncture in Plain Questions is the earliest ancient literature on acupuncture taboo, which has a far-reaching influence on the clinical acupuncture safety regulation. Acupuncture Prohibition of A-B Classic of Acupuncture and Moxibustion absorbs the essence of on Banning of Acupuncture in Plain Questions, retaining most of the contents of the theory and rearranging it. This paper tries to explore the discussions about on Banning of Acupuncture in Plain Questions, compared with Acupuncture Prohibition, from the point of "anatomy" and "spirit", to describe the early understanding of acupuncture taboo, to emphasize the importance of acupuncture safety from three aspects, as human anatomy, needle and operation, and mental and functional status. It is suggested that a relatively preliminary acupuncture safety regulation was being formed at the time, in order to bring inspiration to the contemporary acupuncture clinical .