[Effects of catgut embedding and PGLA embedding at "Zusanli" (ST 36) on skin mast cells, substance P and histamine in healthy rats].

OBJECTIVE: To observe the effects of catgut embedding and polyglycolic acid/poly-lactic acid (PGLA) embedding at "Zusanli" (ST 36) on the activation of local skin mast cells (MC), and expression of substance P (SP) and histamine (HA), and to explore the mechanism of the temporal stimulation effect of acupoint catgut embedding and provide a foundation for further research on the initiation mechanism of acupoint catgut embedding. METHODS: One hundred and sixty male SPF-grade SD rats were randomly divided into a blank group (10 rats), a sham-embedding group (50 rats), a catgut group (50 rats), and a PGLA group (50 rats). Each intervention group was further randomly divided into five subgroups according to the time points after intervention: 8 hours, 3 days, 7 days, 14 days, and 21 days, with 10 rats in each subgroup. One-time sham-embedding, catgut embedding and PGLA embedding was given at left "Zusanli" (ST 36) in each intervention group, respectively. The skin and subcutaneous connective tissue of the left "Zusanli" (ST 36) were collected at the corresponding time points after intervention, except for the blank group (only one day before intervention). Toluidine blue staining was used to detect MC count and degranulation, and immunohistochemical staining was used to detect the expression of SP and HA positive cells. RESULTS: There was no significant difference in MC count between the subgroups of each intervention group and the blank group (P>0.05). There was no significant difference in MC count between the subgroups of the catgut group and the PGLA group (P>0.05). The MC count in the 8-hour subgroup of PGLA group was higher than that in the 8-hour subgroup of catgut group (P<0.05), while the MC count in the 21-day subgroup of PGLA group was lower than that in the 21-day subgroup of catgut group (P<0.05). Compared with the blank group, the degranulation rates of MC were increased in the 8-hour and 3-day subgroups of sham-embedding group, 8-hour, 3-day, and 7-day subgroups of catgut group, and 8-hour, 3-day, 7-day, and 14-day subgroups of PGLA group (P<0.01, P<0.05, P<0.001). There was no significant difference in the degranulation rate of MC between the subgroups of the catgut group and the PGLA group (P>0.05), and no significant difference in the degranulation rate of MC between the two embedding groups at the same time point (P>0.05). Compared with the blank group, the expression of SP positive cells was increased in the 8-hour subgroup of sham-embedding group, 8-hour, 3-day, 7-day, and 14-day subgroups of catgut group, and 3-day, 7-day, and 14-day subgroups of PGLA group (P<0.001, P<0.05). The expression of SP positive cells in the 7-day subgroup of catgut group was higher than that in the 8-hour subgroup of catgut group (P<0.05), while the expression of SP positive cells in the 14-day subgroup of catgut group was lower than that in the 7-day subgroup of catgut group (P<0.001). The expression of SP positive cells in the 7-day subgroup of PGLA group was higher than that in the 3-day subgroup of PGLA group (P<0.05), while the expression of SP positive cells in the 14-day subgroup of PGLA group was lower than that in the 7-day subgroup of PGLA group (P<0.01). There was no significant difference in the expression of SP positive cells between the subgroups of the two embedding groups at the same time point (P>0.05). Compared with the blank group, the expression of HA positive cells was increased in the 8-hour, 3-day subgroups of sham-embedding group, 8-hour, 3-day, 7-day, and 14-day subgroups of catgut group, and 8-hour, 3-day, 7-day, 14-day, and 21-day subgroups of PGLA group (P<0.001, P<0.01, P<0.05). The expression of HA positive cells in the 14-day subgroup of catgut group was lower than that in the 7-day subgroup of catgut group (P<0.05), while the expression of HA positive cells in the 3-day subgroup of PGLA group was higher than that in the 8-hour subgroup of PGLA group (P<0.05), and the expression of HA positive cells in the 14-day subgroup of PGLA group was lower than that in the 7-day subgroup of PGLA group (P<0.05). The expression of HA positive cells in the 3-day subgroup of PGLA group was higher than that in the 3-day subgroup of catgut group (P<0.05). CONCLUSION: Catgut and PGLA embedding at "Zusanli" (ST 36) in healthy rats could induce changes in local skin MC, SP, and HA, which may be one of the mechanisms of the temporal stimulation effect after acupoint embedding. There are certain differences between different suture materials. A moderate inflammatory response in the acupoint area, mediated by MC and involving SP and HA, may be one of the initiating factors for the effect of acupoint catgut embedding.

[Effect of catgut implantation on macrophage CD68, tumor necrosis factor-α and interleukin-1ß in "Zusanli" (ST 36) region of rats].

OBJECTIVE: To observe the expression of local macrophages and related cytokines tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) after catgut implantation in "Zusanli"(ST 36) in rats, so as to explore its underlying mechanisms in inducing therapeutic effect. METHODS: A total of 110 male SD rats were randomly divided into blank control group (n=10), catgut embedding (CE) group (n=50), and sham CE group (n=50). The CE and sham CE groups were randomly divided into 8 h, 3 d, 7 d, 14 d and 21 d subgroups after the intervention (n=10 in each time point group). Rats of the CE group were uniformly subjected into catgut embedding at ST36 once, and those of the sham CE group received embedding needle puncture at ST36 without catgut retention, and the blank control group was only grasped and fixed without other treatments. Tissues from the ST36 area in each group were collected at the corresponding time points, and the expression of CD68 in macrophages in the acupoint area was detected by immunofluorescence, the contents of TNF-α and IL-1ß in the acupoint area were detected by ELISA. RESULTS: Following catgut embedment at ST36, the contents of TNF-α and IL-1ß, and macrophage CD68 expression level began to increase at 8 h, peaked at 3 d, and then gradually decreased at 7, 14, and 21 d, being still higher in the CE group than in the blank control group at 21 d (P<0.05). Compared with the blank control group, the contents of TNF-α and IL-1ß, and macrophage CD68 expression were significantly increased at 8 h, and 3, 7, 14 and 21 d in the CE group (P<0.05). Following sham CE at ST36, the content of TNF-α at 8 h and 3 d, IL-1ß at 8 h and 3, 7 and 14 d, and expression of CD68 at 8 h were significantly increased in comparison with the blank control group (P<0.05). Comparison between the CE and sham CE groups showed that the contents of IL-1ß at 3, 7, 14 and 21 d, and contents of TNF-α，CD68 expression at 8 h, and 3, 7, 14 and 21 d were significantly higher in the CE group than in the sham CE group (P<0.05). CONCLUSION: Catgut embedding at ST36 can induce an increase levels of inflammatory cytokines TNF-α, IL-1ß and macrophage CD68 in the local microenvironment in rats, which may contribute to its functions in initiating therapeutic effect.

[Acupoint injection ameliorates Th1/Th2 imbalance through Toll-like receptor 4/activator protein-1 signal pathway and improves inflammatory response in rats with allergic rhinitis].

OBJECTIVE: To observe the effect of acupoint injection on serum T helper (Th)1/Th2 related cytokines, and the expression levels of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and activator protein-1 (AP-1) of nasal mucosa in allergic rhinitis (AR) rats, so as to explore its mechanism underlying improvement of AR. METHODS: Thirty-two SD rats were randomly divided into normal, model, non-acupoint injection and acupoint injection groups (n=8 in each group). The AR model was established by ovalbumin sensitization. In the acupoint injection group, "Yintang" (GV24+) and bilateral "Yingxiang" (LI20) were selected for injection of mixture solution of dexamethasone and lidocaine (0.05 mL/acupoint), once every 4 days for a total of 4 times. The non-acupoints, located at the midpoint between the "Houhai" and "Huantiao" (GB30) on the bilateral hips and the sites 5 cm inferior to the axillary were injected with the same dose of mixture solution as that in the acupoint injection group. The AR severity was assessed by cumulative quantification scoring methods (including the numbers of nose-catching and sneezes, and the amount of nasal secretions in 30 min). The pathological changes of nasal mucosa were observed by HE staining. The contents of immunoglobulin E (IgE), interleukin (IL)-4 and interferon (IFN)-γ in serum were detected by ELISA. The expressions of TLR4 and MyD88 in nasal mucosa was detected by immunofluorescence. The expression of AP-1 in nasal mucosa was detected by Western blot. RESULTS: Following modeling, the AR symptom score, serum IgE and IL-4 contents and expression of TLR4, MyD88 and AP-1 of nasal mucosa were significantly increased in the model group than those in the normal group (P<0.01), while the serum IFN-γ content was significantly decreased (P<0.01). Compared with the model group and non-acupoint injection group, the AR symptom score, the serum contents of IgE and IL-4 and the expressions of TLR4, MyD88 and AP-1 in nasal mucosa were significantly decreased in the acupoint injection group (P<0.01, P<0.05), while the serum IFN-γ content was significantly increased (P<0.01). H.E. staining of the nasal mucosa showed that most of the epithelium fell off, the lamina propria vessels expanded, the glands proliferated, and eosinophils and lymphocytes infiltrated in the model and non-acupoint injection groups, and those were significantly improved in the acupoint injection group. CONCLUSION: Acupoint injection can effectively improve allergic inflammation of the nose in AR rats, which may be related with its function in inhibiting the abnormal activation of TLR4/AP-1 signaling pathway and regulating the imbalance of Th1/Th2.

Effects of catgut embedding and PGLA embedding at "Zusanli" (ST 36) on skin mast cells, substance P and histamine in healthy rats / 中国针灸

OBJECTIVE@#To observe the effects of catgut embedding and polyglycolic acid/poly-lactic acid (PGLA) embedding at "Zusanli" (ST 36) on the activation of local skin mast cells (MC), and expression of substance P (SP) and histamine (HA), and to explore the mechanism of the temporal stimulation effect of acupoint catgut embedding and provide a foundation for further research on the initiation mechanism of acupoint catgut embedding.@*METHODS@#One hundred and sixty male SPF-grade SD rats were randomly divided into a blank group (10 rats), a sham-embedding group (50 rats), a catgut group (50 rats), and a PGLA group (50 rats). Each intervention group was further randomly divided into five subgroups according to the time points after intervention: 8 hours, 3 days, 7 days, 14 days, and 21 days, with 10 rats in each subgroup. One-time sham-embedding, catgut embedding and PGLA embedding was given at left "Zusanli" (ST 36) in each intervention group, respectively. The skin and subcutaneous connective tissue of the left "Zusanli" (ST 36) were collected at the corresponding time points after intervention, except for the blank group (only one day before intervention). Toluidine blue staining was used to detect MC count and degranulation, and immunohistochemical staining was used to detect the expression of SP and HA positive cells.@*RESULTS@#There was no significant difference in MC count between the subgroups of each intervention group and the blank group (P>0.05). There was no significant difference in MC count between the subgroups of the catgut group and the PGLA group (P>0.05). The MC count in the 8-hour subgroup of PGLA group was higher than that in the 8-hour subgroup of catgut group (P<0.05), while the MC count in the 21-day subgroup of PGLA group was lower than that in the 21-day subgroup of catgut group (P<0.05). Compared with the blank group, the degranulation rates of MC were increased in the 8-hour and 3-day subgroups of sham-embedding group, 8-hour, 3-day, and 7-day subgroups of catgut group, and 8-hour, 3-day, 7-day, and 14-day subgroups of PGLA group (P<0.01, P<0.05, P<0.001). There was no significant difference in the degranulation rate of MC between the subgroups of the catgut group and the PGLA group (P>0.05), and no significant difference in the degranulation rate of MC between the two embedding groups at the same time point (P>0.05). Compared with the blank group, the expression of SP positive cells was increased in the 8-hour subgroup of sham-embedding group, 8-hour, 3-day, 7-day, and 14-day subgroups of catgut group, and 3-day, 7-day, and 14-day subgroups of PGLA group (P<0.001, P<0.05). The expression of SP positive cells in the 7-day subgroup of catgut group was higher than that in the 8-hour subgroup of catgut group (P<0.05), while the expression of SP positive cells in the 14-day subgroup of catgut group was lower than that in the 7-day subgroup of catgut group (P<0.001). The expression of SP positive cells in the 7-day subgroup of PGLA group was higher than that in the 3-day subgroup of PGLA group (P<0.05), while the expression of SP positive cells in the 14-day subgroup of PGLA group was lower than that in the 7-day subgroup of PGLA group (P<0.01). There was no significant difference in the expression of SP positive cells between the subgroups of the two embedding groups at the same time point (P>0.05). Compared with the blank group, the expression of HA positive cells was increased in the 8-hour, 3-day subgroups of sham-embedding group, 8-hour, 3-day, 7-day, and 14-day subgroups of catgut group, and 8-hour, 3-day, 7-day, 14-day, and 21-day subgroups of PGLA group (P<0.001, P<0.01, P<0.05). The expression of HA positive cells in the 14-day subgroup of catgut group was lower than that in the 7-day subgroup of catgut group (P<0.05), while the expression of HA positive cells in the 3-day subgroup of PGLA group was higher than that in the 8-hour subgroup of PGLA group (P<0.05), and the expression of HA positive cells in the 14-day subgroup of PGLA group was lower than that in the 7-day subgroup of PGLA group (P<0.05). The expression of HA positive cells in the 3-day subgroup of PGLA group was higher than that in the 3-day subgroup of catgut group (P<0.05).@*CONCLUSION@#Catgut and PGLA embedding at "Zusanli" (ST 36) in healthy rats could induce changes in local skin MC, SP, and HA, which may be one of the mechanisms of the temporal stimulation effect after acupoint embedding. There are certain differences between different suture materials. A moderate inflammatory response in the acupoint area, mediated by MC and involving SP and HA, may be one of the initiating factors for the effect of acupoint catgut embedding.

Hierarchical MXene/Polypyrrole-Decorated Carbon Nanofibers for Asymmetrical Capacitive Deionization.

Membrane capacitive deionization (MCDI) has emerged as a promising electric-field-driven technology for brackish water desalination and specific salt or charged ion separation. The use of carbon-based or pseudocapacitive materials alone for MCDI usually suffers from the drawbacks of low desalination capacity and poor cycling stability due to their limited accessible adsorption sites and obstructed charge-carrier diffusion pathways. Therefore, developing a hybrid electrode material with multiple charge storage mechanisms and continuous electron/ion transport pathways that can synergistically improve its specific capacitance and cycling durability has currently become one of the most critical technical demands. Herein, we developed a novel hierarchically architectured hybrid electrode by first spinning MXene into polyacrylonitrile (PAN)-based carbon nanofibers (MCNFs) to obtain a highly conductive carbon nanocomposite framework. The excellent spatial support structure can effectively prevent the dense packing of Cl-- and DBS--doped polypyrrole (PPy) molecular chains during the following electrodeposition process, which not only ensures the efficient transport of electrons in the continuous hybrid carbon nanofibrous skeleton but also provides abundant accessible sites for ion adsorption and insertion. The obtained self-supporting membrane electrodes (MCNF@PPy+Cl- and MCNF@PPy+DBS-) have the advantages of outstanding specific capacitance (318.4 and 153.9 F/g, respectively), low charge transfer resistance (10.0 and 5.3 Ω, respectively), and excellent cycling performance (78% and 90% capacitance retention ratios, respectively, after 250 electrochemical cycles). Furthermore, the asymmetrical membrane electrodes showed a superior desalination capacity of 91.2 mg g-1 in 500 mg/L NaCl aqueous solution and obvious divalent ion (Ca2+, Mg2+) selective adsorption properties in high-salt water from the cooling towers of thermal power plants.

[Bibliometric analysis on key elements of clinical application of acupoint catgut embedding therapy in recent ten years].

OBJECTIVE: To analyze the characteristics of acupoint catgut embedding methods, tools, catgut types, and the treatment cycles in the clinical research in recent ten years both at home and abroad, so as to summarize its regularities and to provide technical references for further studies. METHODS: Articles about clinical researches on catgut embedding therapy published in recent ten years (from January 1, 2010 to December 31,2020) were retrieved from the databases of CNKI, Wanfang, VIP, and PubMed by using key words of "acupoint embedding" "acupoint catgut embedding" and "catgut implantation at acupoint". According to the inclusion and exclusion criteria, a new database was established for analyzing the data mentioned above. RESULTS: 1) A total of 1 196 articles were collected, including 15 English articles and 1 181 Chinese articles, presenting a fluctuating increasing trend in recent ten years. 2) The commonly used acupoint embedding methods included disposable catgut embedment needle method (399 times, 38.89%) and disposable syringe needle catgut embedding method (347 times, 33.82%), for which two or multiple methods were mentioned in the same one article. 3) The most frequently used top two tools for catgut embedding were the dispo-sable catgut embedment needle (463 times, 43.03%) and disposable syringe needle (406 times,37.73%), with a significant increase in the application of disposable syringe needle. The most commonly used size of tools included No. 7 (283 times, 39.86%), No. 9 (196 times, 27.61%) and No. 8 (109 times, 15.35%). 4) The most frequently implanted surgical suture was still the common catgut (671 times, 58.15%) despite of a reduction in clinical application year by year, and the types of the implanted suture materials were gradually enriched since 2018, such as the absorbable surgical suture, polyethylprolactide(PGLA), collagen protein thread, polydioxanone(PPDO), etc. The commonly used implanted catgut size was 3-0 (227 times, 30.15%), 2-0 (176 times, 23.37%), 4-0 (131 times, 17.40%), 0 (103 times, 13.68%), with the commonly used catgut length being 1 cm (332 times, 35.55%), 1.5 cm (103 times, 11.03%), 1-2 cm (92 times, 9.85%) and 2 cm (92 times, 9.85%). 5) The intervals of the catgut implantation were 7 days (313 times, 28.95%), 14 days (262 times, 24.24%), 10 days (174 times, 16.10%), and 15 days (162 times, 14.99%). CONCLUSION: In recent ten years, clinical research on acupoint catgut embedding is growing rapidly, and the embedding methods, tools, implanted sutures, and embedding intervals are various, which may provide certain technical references for the future researches and suggest an urgent need of formulation of the standardized and unified standards in this field.

[Effect of moxibustion combined with acupoint catgut embedding on IL-6/JAK/STAT3 signaling pathway in colonic mucosa of ulcerative colitis rats].

OBJECTIVE: To observe the effect of moxibustion combined with acupoint catgut embedding on the content of interleukin 6 (IL-6), and the expressions of janus activated kinase (JAK), signal transducer and activator of transcription 3 (STAT3) in colonic mucosa of rats with ulcerative colitis (UC), so as to explore its mechanism underlying improvement of UC. METHODS: Thirty SD rats were randomly divided into normal, model, acupoint catgut embedding (ACE), moxibustion and acupoint catgut embedding combined with moxibustion (combination) groups (n=6 rats in each group). The UC model was established by enema of trinitro-benzene-sulfonic acid and ethanol. Moxibustion was applied to bilateral "Tianshu" (ST25), "Dachangshu" (BL25) and "Shangjuxu" (ST37) for 10 min, once daily for 14 days, and ACE applied to the same 3 acupoints, once a week for two weeks. After the treatment, colonic mucosal pathological changes were observed by H.E. staining, the level of IL-6 in colonic mucosa was assayed by ELISA, and the expressions of JAK and STAT3 in colonic mucosa were detected by immunohistochemistry and Western blot. RESULTS: H.E. staining showed severe defect of the colonic mucosal epithelium with infiltration of a large number of inflammatory cells in the model group, which was milder in moxibustion, ACE and combination groups. After modeling, the content of colonic IL-6, and the expression levels of JAK and STAT3 were obviously increased (P<0.01) in the model group relevant to the normal group. Following the intervention, the increase of IL-6 contents, and JAK and STAT3 expressions were reversed (P<0.05, P<0.01) in moxibustion, ACE and combination groups. The therapeutic effects of moxibustion combined with ACE were considerably superior to those of simple ACE and simple moxibustion in down-regulating the levels of JAK and STAT3 expression (P<0.01). CONCLUSION: Acupoint catgut embedding combined with moxibustion can repair the injured colonic mucosa of UC rats, which may be related with its effect in suppressing the activation of IL-6/JAK/STAT3 signaling pathway.

[Synergistic effect of acupoint injection and moxibustion or catgut embedding or acupuncture on symptoms and expression of Th1/Th2 related cytokines in nasal mucosa of rats with allergic rhinitis].

OBJECTIVE: To observe the effect of moxibustion, catgut embedding and acupuncture on allergic symptoms and expression of interferon-γ (IFN-γ) and interleukin-4 (IL-4) of nasal mucosa in rats with allergic rhinitis (AR) based on acupoint injection, so as to explore their synergistic effect and related mechanism in relieving AR. METHODS: SD rats (half male half female) were randomly divided into normal control, model, acupoint injection (AI), AI+moxibustion, AI+catgut embedding and AI+acupuncture groups, with 8 rats in each group. The AR model was established by intraperitoneal injection of ovalbumin suspension (once every other day for 7 times), and intranasal drop of 0.5% ovalbumin solution (once daily for 7 days). After successful modeling, rats of the AI group received injection of a mixture solution of equal proportion of 1% lidocaine, dexamethasone and transfer factor into "Yingxiang" (LI20) and "Yintang" (EX-HN3) once every 4 days, 4 times altogether. Mild moxibustion or catgut embedment or manual acupuncture was applied to bilateral "Feishu" (BL13) and "Zusanli" (ST36). Both moxibustion (20 min every time) and acupuncture (with the needles retained for 30 min every time) were conducted once daily for 14 times, and catgut embedding was conducted once a week, twice altogether based on acupoint injection. The rats' nasal allergic reaction score (symptom score, 1-3 points) was given according to the times of nose scratching and sneezing, and the running nose state in 30 min, and histopathological changes of nasal mucosa were observed by H.E. staining. The expression levels of IFN-γ and IL-4 in the nasal mucosa were detected by immunohistochemistry and Western blot, separately. RESULTS: Compared with the normal group, the symptom score and the expression of IL-4 positive cells and protein in nasal mucosa were significantly increased (P<0.05), while the expression of IFN-γ positive cells and protein were considerably decreased in the model group (P<0.05). In comparison with the model group, the symptom score and the expression of IL-4 positive cells and protein were obviously decreased (P<0.05), while the expression of IFN-γ positive cells and protein were remarkably increased in the 4 treatment groups (P<0.05). The effects of AI+moxibustion, AI+catgut embedment, AI+acupuncture were signi-ficantly superior to those of simple AI in up-regulating the expression of IFN-γ positive cells and protein and in down-regulating the expression of IL-4 positive cells and protein (P<0.05). Both the symptom score and the expression of IL-4 were notably lower in the AI+moxibustion group than in the AI+catgut embedment and AI+acupuncture groups (P<0.05), whereas the expression of IFN-γ was apparently higher in the AI+moxibustion group than in the other 3 treatment groups (P<0.05). No significant differences were found between the AI+catgut embedment and AI+acupuncture groups in the levels of symptom score, IFN-γ and IL-4 expressions (P>0.05). CONCLUSION: Moxibustion or catgut embedment or acupuncture and AI have a synergistic effect in relieving symptoms of AR rats, which may be related to their function in regulating the expression levels of nasal IFN-γ and IL-4 proteins. The therapeutic effect of moxibustion is obviously superior to those of both acupuncture and catgut embedment.

[Effect of acupoint catgut embedding combined with moxibustion on Notch signaling pathway in colonic mucosa of rats with active ulcerative colitis].

OBJECTIVE: To observe the effect of acupoint catgut embedding (ACE) combined with moxibustion on the expressions of Notch receptor 1 and target genes Hes 1 and Math 1 in colonic mucosa of ulcerative colitis (UC) rats, so as to reveal its mechanism of treating UC from the perspective of Notch signaling pathway. METHODS: Thirty SD rats were randomized into normal, model, ACE combined with moxibustion (combination), ACE, and moxibustion groups (n=6 in each group). The UC model was established by intrarectal administration of 5% trinitro-benzene-sulfonic acid and 50% ethyl alcohol. Moxibustion was applied to bilateral "Shangjuxu" (ST37), "Tianshu" (ST25) and "Dachangshu" (BL25) for 10 min, once daily for 14 days, and ACE applied to the same 3 acupoints, once a week for two weeks. After the treatment, colonic mucosal pathological changes were observed after hematoxylin eosin (HE) stain. Protein and mRNA expressions of Notch 1, Hes 1 and Math 1 were detected by RT-PCR and Western blot, respectively. RESULTS: H.E. staining showed severe defect of the colonic mucosal epithelium with infiltration of a large number of inflammatory cells in the model group, which was milder in moxibustion, ACE and moxibustion + ACE groups. After modeling, the protein and mRNA expressions of Notch 1 and Hes 1 in the colonic mucosa were obviously increased (P<0.01), while the protein and mRNA expressions of Math 1 were significantly decreased (P<0.01) in the model group relevant to the normal group. Following the intervention, the protein and mRNA expression levels of Notch 1 and Hes 1 in the colonic mucosa were significantly lower (P<0.05, P<0.01) in moxibustion, ACE and combination groups, the mRNA expression level of Math 1 was significantly increased in both ACE and combination groups (P<0.01), and the protein expression of Math 1 was significantly higher in the three treatment groups relevant to the model group (P<0.01). The above indexes of moxibustion + ACE were considerably superior to those of simple ACE and simple moxibustion (P<0.05, P<0.01). CONCLUSION: Acupoint catgut embedding combined with moxibustion can repair the injured colonic mucosa of UC rats, which may be related with its effects in suppressing the expression of Notch 1 and Hes 1, up-regulating the expression of Math 1 in the colonic mucosa, thereby regulating the balance of differentiation between colonic epithelial secretory and absorption cell lines.

A signal peptide derived from Hsp60 induces protective cytotoxic T lymphocyte immunity against lymphoid malignancies independently of TAP and classical MHC-I.

Hsp60sp, a signal peptide derived from the leader sequence of heat shock protein 60 kDa (Hsp60), is a Qa-1/HLA-E-binding peptide. We previously showed that Hsp60sp-specific CD8+ T cells are involved in the immunoregulation of autoimmune diseases by controlling the response of self-reactive lymphocytes. Here, we report that Hsp60sp-specific CD8+ T cells killed malignant lymphocytes in vitro independently of transporter associated with antigen processing (TAP) and classical MHC-I expression. Induction of this cytotoxic T lymphocyte (CTL) response in vivo, either by adoptive transfer of in vitro-amplified CTLs or peptide-loaded dendritic cell immunization, resulted in effective control of lymphoid tumors, including TAP- or classical MHC-I-deficient cells. Hsp60sp-specific immune activation combined with programmed cell death protein 1 (PD-1) blocking synergistically restrained mouse lymphoma development. Importantly, Hsp60sp-specific CD8+ T cells did not negatively affect normal tissues and cells. Our data suggest that Hsp60sp-based immunotherapy is an inviting strategy to control lymphoid malignancies.

[Moxibustion combined with acupoint catgut embedding promotes recovery of injured colonic mucosa by suppressing inflammation in ulcerative colitis rats].

OBJECTIVE: To observe the effect of moxibustion and acupoint catgut embedding (ACE) at "Tianshu"(ST25) "Dachangshu"(BL25) and "Shangjuxu"(ST37) on changes of body mass, stool property, histopathological conditions and expression levels of interleukin 6 (IL-6) in colonic mucosa of ulcerative colitis (UC) rats, so as to reveal its anti-inflammatory mechanisms underlying improvement of UC. METHODS: SD rats were randomized into normal, model, moxibustion, ACE and moxibustion＋ACE groups (n＝6 in each group). The UC model was established by enema of trinitro-benzene-sulfonic acid and ethanol. Moxibustion was applied to bilateral ST25, BL25 and ST37 for 10 min, once daily for 14 days, and ACE applied to the same 3 acupoints, once a week for two weeks. After the treatment, the rats' general conditions were observed, and the severity of UC was assessed by using disease activity index (DAI) score. Colonic mucosal pathological changes were observed under microscope after hematoxylin eosin (H.E.) stain, and the expression levels of IL-6 in the colonic mucosa tissue detected by using immunohistochemical stain and Western blot, respectively. RESULTS: After modeling, the DAI score, and expression level of colonic IL-6 protein detected by immunohistochemistry and Western blot were obviously increased in the model group relevant to the normal group (P<0.01). Following the intervention, the increase of DAI score and IL-6 expression were reversed in moxibustion, ACE and moxibustion＋ACE groups (P<0.01, P<0.05). The therapeutic effects of moxibustion＋ACE were considerably superior to those of simple ACE and simple moxibustion in down-regulating the levels of DAI score and IL-6 expression (P<0.01). H．E. staining showed severe defect of the colonic mucosal epithelium with infiltration of a large number of inflammatory cells in the model group, which was milder in moxibustion, ACE and moxibustion＋ACE groups. CONCLUSION: Moxibustion combined with ACE is able to improve the inflammatory injury of colonic mucosa in UC rats, which may be related with its effect in suppressing the expression of colonic IL-6; and the efficacy of moxibustion＋ACE is apparently superior to that of moxibustion and ACE alone.

[Acupoint injection improves allergic rhinitis by balancing Th17/Treg in allergic rhinitis rats].

OBJECTIVE: To observe the effect of acupoint injection on expression of fork head/winged helix protein 3 (Foxp3), retinoic acid-related orphan receptor γt (RORγt) in nasal mucosa and serum interleukin-17 (IL-17) level in allergic rhinitis (AR) rats, so as to explore its mechanism underlying improvement of AR in terms of balancing Th17/Treg. METHODS: Thirty-two SD rats (half male and half female) were randomized into normal control, AR model, acupoint injection and non-acupoint injection groups (n＝8 in each group). The AR model was established by ovalbumin sensitization. In the acupoint injection group, "Yintang" (EX-HN3) and bilateral "Yingxiang"(LI20) were selected for injection of mixture solution of dexamethasone (DEX) and transfer factor and lidocaine (0.1 mL/acupoint), once every 3 days for a total of 4 times. The non-acupoints, located at the mid-point between the "Houhai" (GV1) and "Huantiao"(GB30) on the bilateral hips and the sites 5 cm inferior to the axillary were injected with the same dose of mixture solution as that in the acupoint injection. The AR severity was assessed by cumulative quantification scoring methods (including the numbers of nose-catching and sneezes, and the amount of nasal secretions in 30 min). The expressions of Foxp3 and RORγt in the nasal mucosa were detected by immunohistochemistry. The serum IL-17 content was detected by enzyme linked immunosorbent assay (ELISA)．. RESULTS: The AR symptom score and serum IL-17 content were significantly higher in the AR model group than those in the normal control group (P<0.05), and significantly down-regulated in the acupoint injection group (not in the non-acupoint group) relevant to the AR model group (P<0.05). Following modeling, the expression levels of nasal Foxp3 protein was significantly down-regulated while that of RORγt protein markedly up-regulated in the AR model group relevant to the normal control group (P<0.05), indicating an imbalance between Foxp3 and RORγt activity(P<0.05). After EA intervention, the increased expression of Foxp3 and the down-regulated expression of RORγt were revised in the acupoint injection group (P<0.05) but not in the non-acupoint group (P>0.05). The percentage of the Foxp3 positive cells and the ratio of Foxp3/RORγt were negatively correlated with the AR symptom score(P<0.05), the expression of RORγt and the content of IL-17 were positively correlated with the symptom score (P<0.05)．. CONCLUSION: Acupoint injection is able to improve symptoms of RA rats, which may be related with its function in up-regulating the level of nasal mucosal Foxp3 and suppressing the levels of nasal RORγt and serum IL-17 to correct the imbalance of Th17/Treg.

[Time-effect reaction observation of micro-invasion embedded poly glycolide-co-lactide suture in Sanyinjiao (SP6) in healthy subjects by using magnetic resonance].

OBJECTIVE: To observe the time-effect of stimulation of the embedded poly glycolide-co-lactide (PGLA) suture in Sanyinjiao (SP6) area in normal human body, so as to provide an experimental evidence for clinical application of micro-invasion suture-embedding at an appropriate interval. METHODS: A total of 8 healthy volunteer students (3 boys and 5 girls, ranging in age from 24 to 27 years) were recruited in the present study. A piece of sterilized PGLA suture was implanted into the left SP6 using minimally invasive surgery after strict local skin disinfection. The fat-suppression T2 weighted magnetic resonance images (MRI, displaying local lesion after eliminating interference of fat tissue signals), and T2 mapping 8-echo train images were acquired before and 8 h, 3, 7, 10 and 14 days after PGLA suture embedment by using a MR imaging system. After transformation of the T2-mapping 8-echo train images into T2-mapping images by using a relevant software, the T2 values (meaning the relaxation time of the local muscle) of the left SP6 were measured, followed by analysis of the signal intensity of T2 weighted fat-suppression images and T2 values at different time-points. RESULTS: Before the suture embedding, no abnormal signals were found in the signal intensity of T2 weighted fat-suppression images. After PGLA suture embedment, the local signal intensity of T2WI fat-suppression images was relatively increased at the 8th h, and on day 3, 7, 10 and 14 relevant to pre-embedment, but gradually atte-nuated on day 10 and 14. The T2 values were significantly increased at the 5 time-points of post-embedment (all P<0.01), but without significant differences among the 8thh, the 3rd and 7thd (P>0.05), and being markedly lowered on day 14 relevant to day 7 (P<0.01) in spite of being still markedly higher than that of pre-embedding (P<0.01). CONCLUSION: The signal intensity of T2 weighted fat-suppression images and T2 values acquired from PGLA-suture-embedded SP6 acupoint area in healthy subjects may keep at least for 2 weeks, suggesting that the stimulating reaction of suture-embedment persists more than 14 days. Hence, when a micro-invasion embedding with PGLA suture performed, the interval of two weeks would be appropriate.

[Observation on time-effect for the stimulating at Zusanli (ST 36) with minimally invasive embedding with PGLA in the healthy person].

OBJECTIVE: To explore the objectivity and time-effect of stimulating effect at acupoint with PGLA in the healthy person, and to provide a basis for the rational interval of minimally invasive embedding of PGLA. METHODS: Before embedding, 8 h, 3rd, 7th, 10th, 14th day after embedding, medical imaging magnetic resonance imaging (MRI) scanning technique was used to collect local T2WI pressure-lowering and T2-Mapping 8 echoes sequence image of left Zusanli (ST 36) in 8 cases of healthy person. The T2-Mapping 8 echoes sequence image was generated by the relevant software to the T2-Mapping image and the local T2 value was measured. The characteristics of local T2WI pressure-fat image signal intensity and the change of T2 value at left Zusanli (ST 36) with minimally invasive embedding with PGLA were observed and analyzed. RESULTS: ①There was no abnormal signal on the T2WI pressure-fat image on the left Zusanli (ST 36) point before the embedding. The high-signal was seen on the local T2WI pressure-fat image at each time point after embedding, there was no significant difference in local signal intensity between 8 h, 3rd and 7th day after embedding. The local signal intensity decreased on the 10th day after embedding, and the local signal intensity decreased significantly on the 14th day after embedding.②The T2 value at each time point after embedding increased significantly compared with that before embedding (all P<0.01); there was no significant difference in T2 value among the 8 h, 3rd and 7th day after embedding (all P>0.05); there was no significant difference between the T2 value on the 7th and the 10th day after embedding (P>0.05)，the T2 value on the 14th day after embedding was significantly lower than that on the 7th day after embedding (P<0.01). CONCLUSION: It has a stimulating effect on the local acupoints with minimally invasive embedding with PGLA in the healthy person, and the stimulating effect has certain time-effect. The effective stimulation time is about 2 weeks. The rational interval period for the minimally invasive embedding with the PGLA of the same specification type should be about 2 weeks.

Observation on time-effect for the stimulating at Zusanli (ST 36) with minimally invasive embedding with PGLA in the healthy person / 中国针灸

OBJECTIVE@#To explore the objectivity and time-effect of stimulating effect at acupoint with PGLA in the healthy person, and to provide a basis for the rational interval of minimally invasive embedding of PGLA.@*METHODS@#Before embedding, 8 h, 3rd, 7th, 10th, 14th day after embedding, medical imaging magnetic resonance imaging (MRI) scanning technique was used to collect local T2WI pressure-lowering and T2-Mapping 8 echoes sequence image of left Zusanli (ST 36) in 8 cases of healthy person. The T2-Mapping 8 echoes sequence image was generated by the relevant software to the T2-Mapping image and the local T2 value was measured. The characteristics of local T2WI pressure-fat image signal intensity and the change of T2 value at left Zusanli (ST 36) with minimally invasive embedding with PGLA were observed and analyzed.@*RESULTS@#①There was no abnormal signal on the T2WI pressure-fat image on the left Zusanli (ST 36) point before the embedding. The high-signal was seen on the local T2WI pressure-fat image at each time point after embedding, there was no significant difference in local signal intensity between 8 h, 3rd and 7th day after embedding. The local signal intensity decreased on the 10th day after embedding, and the local signal intensity decreased significantly on the 14th day after embedding.②The T2 value at each time point after embedding increased significantly compared with that before embedding (all 0.05); there was no significant difference between the T2 value on the 7th and the 10th day after embedding (>0.05)，the T2 value on the 14th day after embedding was significantly lower than that on the 7th day after embedding (<0.01).@*CONCLUSION@#It has a stimulating effect on the local acupoints with minimally invasive embedding with PGLA in the healthy person, and the stimulating effect has certain time-effect. The effective stimulation time is about 2 weeks. The rational interval period for the minimally invasive embedding with the PGLA of the same specification type should be about 2 weeks.

[Acupoint Injection at "Yingxiang"(LI 20) and "Yintang"(GV 29) May Relieve Nasal Allergic Symptoms Possibly by Down-regulating Expression of Histamine Receptor H 1 and H 4 in Nasal Mucosa of Allergic Rhinitis Rats].

OBJECTIVE: To observe the effect of acupoint injection at "Yingxiang"(LI 20) and "Yintang"(GV 29) on nasal allergic reactions and the expression of histamine receptor H 1 and H 4 in nasal mucosa of allergic rhinitis (AR) rats, so as to reveal its mechanism underlying improvement of AR. METHODS: SD rats were randomized into normal, model, non-acupoint injection and acupoint injection groups (n=8 in each). The AR model was established by intraperitoneal (i.p.) injection of mixture of ovalbumin, aluminium hydroxide gel and normal saline (once every other day, for 7 times), and nasal drip of ovalbumin (on the following day of i.p.for 7 days). The mixture solution of lidocaine, dexamethasone (DXM) and transfer factor (0.1 mL/acupoint or non-acupoint) was injected into bilateral LI 20 and GV 29 in the acupoint injection group, or into the non-acupoints (about 5 cm below the armpit on both sides, and the middle point between the left "Houhai"[GV 1] and "Huantiao"[GB 30]), on the 1st , 5th, 9th, and 13th day after modeling. Symptoms of sneezing, nasal discharge, nose-rubbing, etc. were scored after the treatment. The expression levels of H1 R and H4 R proteins and genes in the nasal mucosa tissue were determined using immunohistochemistry and quantitative real-time PCR, respectively. RESULTS: Compared with the normal group, the symptom score and the expression levels of H1 R, H4 R proteins and genes in nasal mucosa were significantly increased in the model group (P<0.05). After acupoint injection for 4 times, the symptom score and the expression levels of H1 R, H4 R proteins and mRNAs were significantly down-regulated in the acupoint injection group relevant to the model group (P<0.05). There were no significant differences between the non-acupoint and the model groups in the abovementioned indexes (P>0.05), suggesting a specificity of the effect of acupoint injection. CONCLUSION: Acupoint injection can relieve the allergic symptoms of AR rats, which may be related to its effects in down-regulating the over expression of H1 R and H4 R proteins and genes in the nasal mucosa.

[Effect of Acupoint Injection on Eosinophil Counts,Protein and mRNA Expressions of Eotaxin in Nasal Mucosa of Allergic Rhinitis Rats].

OBJECTIVE: To observe the effect of acupoint injection on eosinophils (EOS) counts and expression of eotaxin in nasal mucosa of allergic rhinitis (AR) rats, so as to reveal its mechanism underlying improving AR. METHODS: Twenty-four Sprague-Dawley (SD) rats were randomly divided into normal, model and acupoint injection groups (n=8 in each group). The AR model was established by intraperitoneal injection of ovalbumin sensitization. Bilateral "Yingxiang"(LI 20) and "Yintang"(GV 29) were selected for acupoint injection of the mixture solution of lidocaine, dexamethasone, and transfer factor (0.1 mL/acupoint) on the 1st, 5th, 9th, and 13th day after AR model established, a total of four times. EOS in the nasal mucosa was counted under light microscope after HE staining. Protein and mRNA expressions of eotaxin in the nasal mucosa were detected by immunohistochemical and RT-PCR methods, respectively. RESULTS: Compared with the normal group, EOS counts, protein and mRNA expressions of eotaxin in the nasal mucosa were significantly higher in the model group (P<0.05). Compared with the model group, EOS counts, protein and mRNA expressions of eotaxin in the nasal mucosa were significantly lower in the acupoint injection group (P<0.05). CONCLUSIONS: Acupoint injection can reduce the nasal mucosa inflammation by suppressing the protein and mRNA expressions of eotaxin, decreasing the infiltration and gathering of EOS in the nasal mucosa.

[Analysis on Influencing Factors of Acupoint Injection of Drugs for Improving Allergic Rhinitis in Rats].

OBJECTIVE: To observe the therapeutic effect of acupoint injection of mixture solution of lidocaine, dexa-methasone (DX) and transfer factor on symptoms and serum histamine contents in allergic rhinitis (AR) rats, so as to differentiate the role of acupoint stimulation or drug factor influencing the therapeutic effect. METHODS: SD rats were randomized into normal (n=8), model, non-acupoint-drug injection, acupoint-drug injection and acupoint-saline injection groups (n=11 in each group). The AR model was established by intraperitoneal injection of suspension of ovalbumin (0.3 mg) and aluminum hydroxide (30 mg), 1 mL, once every other day for 7 days, followed by nose-dripping of 5% ovalbumin (50 µL) for other 7 days. The AR scores for rhinocnesmus severity (1-2 points), sneezing times (1-3 points) and nasal discharge quantity (1-3 points) were assessed by cumulative quantification scoring method. "Yintang" (EX-HN 3) and bilateral "Yingxiang" (LI 20) were used for acupoint injection of mixture solution of 1% Lidocaine, DX and transfer factor (1:1:1, 0.1 mL/acupoint), once every other day for 7 days. The non-acupoints were located at the bilateral sub-costal regions and the mid-point between the left "Houhai" (GV 1) and "Huantiao" (GB 30).The same dose of saline was injected into the aforementioned acupoints for control. Serum histamine content was detected by using ELISA, and the nasal mucosal pathological changes were also observed after H.E. stain. RESULTS: After the treatment, AR modeling induced increase of symptom scores and serum histamine contents were significantly decreased in both acupoint-saline injection and acupoint-drug injection groups (P<0.01) but not in the non-acupoint-drug injection group (P>0.05), and the effects of acupoint-drug injection were markedly superior to those of non-acupoint-drug injection group (P<0.01). CONCLUSIONS: Acupoint injection of mixture solution of Lidocaine, DX and transfer factor is effective in relieving the allergic symptoms and reducing the nasal mucosa inflammation in AR rats, which may be related to its action in suppressing histamine releasing, mainly by acupoint stimulation. It suggests that the therapeutic effect mainly derives from acupoint-injection stimulation, then the injected drugs.

[Dynamic Changes of Fast Spin-echo T 2 Weighted Images Displayed Topical Reactions to Micro-invasive Catgut-embedment of Zusanli (ST 36) in Healthy Volunteer Subjects].

OBJECTIVE: To observe the duration of local reactions after subcutaneous micro-invasive catgut-embedding stimulation in Zusanli(ST 36) region in normal human body by using magnetic resonance (MR) T 2-mapping, so as to find a reasonable interval between every two catgut-embedding performance. METHODS: A total of 8 normal volunteers (3 women, and 5 men) were enrolled in the present study. Before and 8 h, and 3, 7, 14, 21, 28 and 35 days after the subcutaneous micro-invasive embedment of catgut (Gauge 2.0, 2 cm long) in the left Zusanli (ST 36) area, the local T 2-weighted images (T 2 WI) and fat-suppression T 2-weighted images (FS T 2 WI) were detected and recorded by using MR scan (GE Optima MR 360 1.5 T) and analyzed using FuncTool software. RESULTS: Compared with pre-catgut-embedment, the local signal strength of T 2 FS WI and T 2 relaxation time began to rise on the 8th hour after catgut-embedment, peaked on the 3rd day, began to descend on the 7th day, but being still higher than pre-embedment, and turned to the normal level on the 21st day to the 35th day. Results of statistical analysis displayed that compared with pre-catgut-embedment, T 2 relaxation time levels were significantly higher on the 8th h, day 3, 7 and 14 after the embedment (P<0.01), and were comparable to that of pre-embedment on day 21, 28 and 35 (P>0.05). CONCLUSIONS: Catgut-embedment of Zusanli (ST 36) induces an apparent change of MRI signals from the 8th h to the 14th day in normal volunteer subjects, suggesting that an interval of 21 days may be reasonable for the micro-invasion catgut-embedment in clinical practice.

[Characteristic of sludge thermal hydrolysis using microwave heating with alkali addition].

NaOH was added into sludge to improve thermal hydrolysis when heated with microwave irradiation. Volatile suspended solid (VSS) rapidly dissolved into aqueous phase when adding 0.1 g NaOH per gram into suspended solid (SS) of sludge. More than 60% of VSS solubilized at 170 degrees C, and VSS to SS ratio of treated sludge reduced to 25%. For 5 min treatment, soluble COD (SCOD) in treated sludge liquid phase was 9.8, 12.8, 15.1, and 14.5 g/L respectively at 80, 120, 150 and 170 degrees C. Accordingly, the SCOD to TCOD (total COD) ratio was 24.0%, 31.3%, 36.9% and 35.6%. The pH of the microwave heated sludge decreased with higher temperature and longer hydrolysis time, while the finale pH of different testes were still higher than 10.5. Alkali dose effect on organic matters dissolving was analyzed at the range of 0-0.2 g NaOH into per gram SS. Results showed that when alkali dose was above 0.05 g, the rapid dissolving trend became level for VSS and SS. BMP (biochemical methane potential) test was used to investigate treated sludge digestibility at NaOH dose of 0.05 g with 5 min microwave heating at set temperature and 0.1 g with 1 min heating. Results showed that adding 0.05 g NaOH improved sludge anaerobic digestion, the biogas production was 28.5% higher than control at 150 degrees 2, while adding 0.1 g NaOH would inhibit treated sludge digestion process.