Effects of electroacupuncture on gut microbiota and related inflammatory factors in rats with Crohn disease / 针灸推拿医学（英文版）

Objective:To observe the effects of electroacupuncture(EA)on gut microbiota and serum inflammatory factors interleukin(IL)-1β and tumor necrosis factor(TNF)-α in Crohn disease(CD)model rats. Methods:Thirty-six Sprague-Dawley rats were randomly divided into a normal control(NC)group with 10 rats and a modeling group with 26 rats.In the modeling group,the CD rat model was prepared with 2,4,6-trinitrobenzene sulfonic acid(TNBS)enema.After successful modeling,the rats were randomly divided into a CD model(CD)group,an EA group,and a Western medicine(WM)group.The NC and CD groups received no treatment;the EA group was treated with EA for 20 min each time,with 7 consecutive days'intervention;the WM group received mesalazine enteric-coated tablet solution by gavage once a day for 7 d.The changes in body mass and disease activity index(DAI)were observed.Serum IL-1β and TNF-α were determined by enzyme-linked immunosorbent assay.Hematoxylin-eosin staining was used to observe the pathological changes of colon tissues,and 16S rDNA sequencing was used to analyze the structural changes of gut microbiota. Results:Compared with the NC group,the body mass of rats in the CD group decreased(P<0.01),and the DAI score increased(P<0.01);the colon tissue structure was disordered,and many inflammatory cells were present;also,IL-1β and TNF-α increased significantly(P<0.01).As a result,the diversity of gut microbiota decreased,and the abundance of some conditional pathogenic bacteria(such as Prevotella)increased,while the abundance of beneficial bacteria(such as Lactobacillus,Rochella,and Spirillum)decreased.After the intervention,compared with the CD group,the body mass of rats in the EA group and WM group increased(P<0.01);the DAI score decreased(P<0.01),the colon tissue structure improved,and the IL-1β and TNF-α levels decreased(P<0.01);the diversity of gut microbiota increased(P<0.05),and the abundance of some conditional pathogenic bacteria decreased while the abundance of beneficial bacteria increased in the EA group;whereas the diversity of gut microbiota in the WM group was not statistically different(P>0.05). Conclusion:EA can reduce the damage of colon mucosa,regulate the imbalance of gut microbiota,and inhibit the serum inflammatory factor IL-1β and TNF-α expression in CD rats.

Formulation Rules in Medical Heart Enlightenment for Treatment of Heart Pain from Perspective of "Tangye Jingfa Tu" / 中国实验方剂学杂志

ObjectiveTo explore the formulation rules for the treatment of heart pain in the Medical Heart Enlightenment （《医学心悟》） from the perspective of the "Tangye Jingfa Tu"， thereby providing a way of thinking about the treatment of heart pain in traditional Chinese medicine （TCM） and the study of the Medical Heart Enlightenment. MethodThe "Tangye Jingfa Tu" contained in the Secrets for Auxiliary Cultivation Life： The Essential Method of Using Herbal Medicine for the Differential Treatment of the Five Zang Organs （《辅行诀五脏用药法要》） was used to analyze the nine prescriptions for heart pain in Volume Ⅲ of the Medical Heart Enlightenment by CHENG Guopeng in the Qing dynasty. A "table for analyzing the formulation of the nine prescriptions for heart pain" was developed. The analysis diagrams for decoction and meridian rules in nine prescriptions for heart pain were plotted and the compatible structure of the medicinal flavors was analyzed. ResultThe composition of Chenxiang Jiangqisan for the treatment of Qi-induced heart pain is "five pungent， four bitter， and two sweet drugs"， the composition of Shouniansan for the treatment of blood-induced heart pain is "five bitter， four pungent， three sweet， and one salty drugs"， the composition of Qingzhongtang for the treatment of heat-induced heart pain is "six bitter， three pungent， and two sweet drugs”， the composition of Jiangfutang with Cinnamomi Cortex for cold-induced heart pain is "three pungent and two sweet drugs"， the composition of Xiaobanxia modified with Fuling Tang for treating fluid retention-induced heart pain is "two pungent and two sweet drugs"， the composition of Baohetang for treating heart pain due to dietary stagnation is "five sweet， four pungent， four bitter， and one sour drugs"， the composition of Guipitang for the treatment of deficiency-induced heart pain is "eight sweet， four bitter， three pungent， and one sour drugs"， the combination of Huachongwan for the treatment of worm-induced heart pain is "seven bitter， six pungent， and four sweet drugs"， the composition of Shenzhusan， Congbaijiu， and Shengjiangtang for the treatment of resistance-induced heart pain is "eight pungent， four bitter， and two sweet drugs". ConclusionFrom the perspective of the "Tangye Jingfa Tu"， the Medical Heart Enlightenment is based on the compatibility principle of "pungent-bitter-sweet drugs" in the treatment of heart pain， with heart deficiency treated with salty drugs for tonifying， or bitter-sweet-salty drugs， and heart excess treated with bitter drugs for purging， or sweet-pungent-bitter drugs， mostly applying the transformation rules of five medicinal flavors， i.e.， "sweet-pungent-bitter drugs" and promoting the action of the pungent and sweet drugs acting on the spleen into the heart to relieve and purge the heart. In most cases， the treatment focuses on harmonizing the heart， liver， spleen， and kidneys， with an emphasis on the mother-child relationship and the application of the principles of the five elements generating and controlling each other. If the progression of the disease involves both the mother and child organs， the formulation should adhere to the compatibility rule of "the child organ makes the mother organ excess and the mother organ makes the child organ deficient".

Variation and interaction mechanism between active components in Rheum officinale and rhizosphere soil microorganisms under drought stress / 中国中药杂志

To explore the changes and the reaction mechanisms between soil microecological environment and the content of secon-dary metabolites of plants under water deficit, this study carried out a pot experiment on the 3-leaf stage seedlings of Rheum officinale to analyze their response mechanism under different drought gradients(normal water supply, mild, moderate, and severe drought). The results indicated that the content of flavonoids, phenols, terpenoids, and alkaloids in the root of R. officinale varied greatly under drought stresses. Under mild drought stress, the content of substances mentioned above was comparatively high, and the content of rutin, emodin, gallic acid, and(+)-catechin hydrate in the root significantly increased. The content of rutin, emodin, and gallic acid under severe drought stress was significantly lower than that under normal water supply. The number of species, Shannon diversity index, richness index, and Simpson index of bacteria in the rhizosphere soil were significantly higher than those in blank soil, and the number of microbial species and richness index decreased significantly with the aggravation of drought stresses. In the context of water deficit, Cyanophyta, Firmicutes, Actinobacteria, Chloroflexi, Gemmatimonadetes, Streptomyces, and Actinomyces were the dominant bacteria in the rhizosphere of R. officinale. The relative content of rutin and emodin in the root of R. officinale was positively correlated with the relative abundance of Cyanophyta and Firmicutes, and the relative content of(+)-catechin hydrate and(-)-epicatechin gallate was positively correlated with the relative abundance of Bacteroidetes and Firmicutes. In conclusion, appropriate drought stress can increase the content of secondary metabolites of R. officinale from physiological induction and the increase in the association with beneficial microbe.

Effect of electroacupuncture at "Neiguan" (PC 6) and "Zusanli" (ST 36) on gastrointestinal hormone in the antral tissue of rats with functional dyspepsia / 中国针灸

OBJECTIVES@#To observe the effects of electroacupuncture (EA) at "Neiguan" (PC 6) and "Zusanli"(ST 36) on the gastric emptying rate, the level of serotonin (5-HT) and the protein expression of motilin (MTL), ghrelin, substance P (SP) and vasoactive intestinal peptide (VIP) in the antral tissue of the rats with functional dyspepsia (FD) and explore the effect mechanism of EA in treatment of FD.@*METHODS@#A total of 21 SPF male SD rat pups were randomly divided into a normal group, a model group and an EA group, with 7 rats in each group. In the model group and the EA group, FD model was prepared by the gavage with 0.1% sucrose iodoacetamide solution combined with the modified small platform method. After the successful modeling, EA was applied to "Neiguan" (PC 6) and "Zusanli"(ST 36) in the rats of the EA group, with disperse-dense wave, 20 Hz/100 Hz in frequency, stimulated for 30 min, once daily, for 7 days consecutively. Before and after intervention, the general condition of the rats was observed in each group. After the completion of intervention, the gastric emptying rate was measured, the morphological changes of gastric antral tissue were observed using HE staining, the level of 5-HT was detected with ELISA method, and the protein expression of MTL, ghrelin, SP, and VIP was determined with Western blot method in the antral tissue of rats.@*RESULTS@#In the normal group, the rats were in a good mental state, with lustrous fur, flexible movement and the increase of food intake and body mass. In the model group, the rats were poor in mental state, lack of lustre in fur, preference for the body curled up, reduced activity and response; and a part of rats had loose stool, obviously enlarged gastric body and gastric food retention. In the EA group, the general condition of rats, e.g. the mental state, food intake and activity, were improved, the gastric body got smaller obviously and the gastric food retention was reduced when compared with the model group. The antral structure was intact, the glands were rich and no injury of the gastric mucosa was found, e.g. inflammatory reaction and edema in the rats of each group. Compared with the normal group, the gastric emptying rate was decreased (P<0.01), 5-HT level was increased (P<0.01), the protein expression of MTL and ghrelin was reduced (P<0.01) and that of VIP was elevated (P<0.01) in the rats of the model group. The gastric emptying rate was increased (P<0.01), 5-HT level was decreased (P<0.01), and the protein expression of MTL and ghrelin was elevated (P<0.05, P<0.01) in the rats of the EA group when compared with those in the model group.@*CONCLUSIONS@#Electroacupuncture at "Neiguan" (PC 6) and "Zusanli"(ST 36) may effectively relieve gastric dysfunction, strengthen gastric motility and promote gastric emptying so as to alleviate the symptoms of dyspepsia in FD rats, and its mechanism may be related to the regulation of gastrointestinal hormones in the antral tissue.

Effects of acupuncture at Feishu(BL13)and Tianshu(ST25)on pulmonary function and tissue inflammation in asthma model rats / 针灸推拿医学（英文版）

Objective:To compare the effects of Feishu(BL13)combined with Tianshu(ST25)and Feishu(BL13)alone on pulmonary function and tissue inflammation in asthma model rats. Methods:Forty-eight Sprague-Dawley rats were randomly divided into a normal group,a model group,a lung treatment group,and a lung-intestine treatment group according to the random number table method,with 12 rats in each group.Except for the normal group,rats in the other three groups were sensitized by ovalbumin followed by atomization stimulation to establish the asthma model.After the model was made successfully,rats in the lung treatment group were treated with acupuncture at bilateral Feishu(BL13)for 30 min;rats in the lung-intestine treatment group were treated with acupuncture at bilateral Feishu(BL13)and Tianshu(ST25)for 15 min,respectively,for 30 min in total.Acupuncture was performed once a day for 14 d.Rats in the other two groups did not receive any intervention.After intervention,the pulmonary function of each group of rats was measured by a pulmonary function tester.The histopathological changes stained by hematoxylin-eosin staining and the collagen deposition degree stained by Masson in lung tissue,as well as the inflammatory cells in the bronchoalveolar lavage fluid(BALF)stained by Wright's-Giemsa staining were observed under a light microscope;the levels of interleukin(IL)-4,IL-5,IL-13,IL-17,IL-25,IL-33,leukotrienes(LT),thymic stromal lymphopoietin(TSLP),and prostaglandin D2(PGD2)were detected by enzyme-linked immunosorbent assay. Results:Compared with the normal group,the peak expiratory flow(PEF),the dynamic lung compliance(Cdyn),the forced expiratory flow 25%(FEF25%),the forced expiratory volume in the first second/forced vital capacity(FEV1/FVC),and the maximum mid-expiratory flow(MMEF)in the model group were significantly decreased(P<0.01 or P<0.05);the lung resistance(RL),the collagen deposition,the IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 levels,and the neutrophil proportion in the BALF were increased significantly(P<0.01 or P<0.05).Compared with the model group,the FEF25%and FEV1/FVC in the pulmonary function were significantly increased(P<0.01,P<0.05),while the collagen fiber deposition and the levels of IL-4,IL-5,IL-13,IL-17,LT,TSLP,and PGD2 were significantly decreased(P<0.01 or P<0.05)in the lung treatment group;the PEF,FEF25%,and FEV1/FVC in pulmonary function were significantly increased(P<0.01 or P<0.05),while the RL and the collagen fiber deposition,as well as the neutrophil proportion and the levels of IL-4,IL-5,IL-13,IL-17,IL-33,LT,TSLP,and PGD2 in the BALF were significantly decreased(P<0.01 or P<0.05)in the lung-intestine treatment group.Compared with the lung treatment group,the IL-5 level in the rat lung tissue of the lung-intestine treatment group was decreased(P<0.05). Conclusion:Acupuncture in the lung treatment and the lung-intestine treatment groups improved pulmonary function,lung inflammation,and inflammatory cells in the BALF in asthma model rats;the lung-intestine treatment was better than lung treatment in improving the IL-5 level.The combination of Feishu(BL13)and Tianshu(ST25)was superior to Feishu(BL13)alone in the treatment of asthma in trend.

Effects of acupuncture and moxibustion on histamine and inflammatory factors in the skin tissue at Tianshu (ST25) of rats with Crohn disease / 针灸推拿医学（英文版）

Objective: To explore the potential mechanism of acupuncture and moxibustion in treating Crohn disease (CD) by evaluating the changes in histamine and inflammatory factors in the skin tissue at Tianshu (ST25) of rats.Methods: Fifty-eight Sprague-Dawley rats were randomly divided into a normal group (n=14) and a CD-modeling group (n=44). Rats in the CD-modeling group received enema with 2,4,6-trinitrobenzene sulfonic acid plus ethanol to establish CD models. The enema was repeated once every 7 d for a total of 4 times. After modeling, four modeled rats and four normal rats were randomly selected for model identification. After the CD model was successfully established, the remaining rats in the CD-modeling group were randomly divided into a model group, an acupuncture group, a moxibustion group, and a Western medication group, with ten rats in each group. The rats in the acupuncture and moxibustion groups were treated with acupuncture or moxibustion at Tianshu (ST25) and Shangjuxu (ST37); the rats in the Western medication group were treated with mesalazine enteric-coated tablets by gavage for continuous 7 d. After the intervention, the colon tissue of rats in each group was collected. After gross observation, hematoxylin-eosin (HE) staining was performed to further observe the pathological changes. The expression of histamine in the skin tissue at Tianshu (ST25) was detected by enzyme-linked immunosorbent assay. The expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-18, IL-10, and IL-6 in the skin tissue at Tianshu (ST25) was detected by Western blotting. Results: Compared with the normal group, the colonic wall of rats in the model group showed cobblestone-like changes, local ulcers, and polyps in dark red and thickening and hardening. HE staining showed local loss of mucosal epithelial layer and formation of slit-like ulcers, destruction of mucosal glands, edema, and infiltration of inflammatory cells in lamina propria and submucosa, and occasional formation of sarcoid-like granuloma. The levels of histamine and IL-6 were significantly up-regulated (P<0.01, P<0.05), and the levels of TNF-α, IL-18, and IL-10 were significantly down- regulated (P<0.01 or P<0.05) in the skin tissue at Tianshu (ST25) of rats in the model group. Compared with the model group, the pathomorphological damage of the colon tissue of rats in the acupuncture group, moxibustion group, and Western medication group was significantly improved. The levels of histamine and IL-6 were significantly down- regulated (P<0.01, P<0.05), and the level of IL-10 was significantly up-regulated (P<0.01) in the skin at Tianshu (ST25) of rats in the acupuncture group. The levels of histamine and IL-6 were significantly down-regulated (P<0.01, P<0.05), and the levels of TNF-α, IL-18, and IL-10 were significantly up-regulated (P<0.01 or P<0.05) in the skin tissue at Tianshu (ST25) of rats in the moxibustion group. The level of histamine was significantly down-regulated (P<0.01), and the levels of IL-18 and IL-10 were significantly up-regulated (P<0.05, P<0.01) in the skin tissue of rats in the Western medication group. Compared with the acupuncture group, the level of IL-10 in the skin tissue at Tianshu (ST25) of rats in the moxibustion group was significantly up-regulated (P<0.01). Conclusion: The inflammatory responses in the skin tissue at Tianshu (ST25) may be the external manifestation of CD. Significant differences in the regulation of inflammatory responses in the skin tissue at Tianshu (ST25) between acupuncture and moxibustion exist, which may be caused by the differences in the stimulation characteristics between acupuncture and moxibustion.

Shenling Baizhusan in Prevention and Treatment of Inflammatory Bowel Disease Based on Theory of "Reinforcing Earth to Generate Metal and Associating Lung with Large Intestine" / 中国实验方剂学杂志

The most common manifestations of inflammatory bowel disease （IBD） are Crohn's disease and ulcerative colitis， and the global incidence of IBD is on the rise. Traditional Chinese medicine （TCM） is advantageous in the treatment of IBD. IBD， with TCM names based on clinical symptoms， mostly belongs to recurrent dysentery， long dysentery， diarrhea， dysentery， bowel， and other categories. In TCM pathogenesis of IBD， spleen deficiency and exuberant dampness predominate in the whole course of the disease. Since the lung is associated with the large intestine and the lung Qi and spleen Qi are interconnected， the lung Qi and spleen Qi are deficient and the dampness and heat accumulate internally， which caused collateral obstruction by stagnant blood and the development of IBD. From the perspective of "associating lung with large intestine"，it is believed that the main mechanism of IBD is the Qi imbalance and abnormal metabolism of fluids in the lung and the intestine，and the nutrient-Yin injury of the lung and the intestine. According to the chronic， recurrent， and diffuse pathogenesis characteristics and main clinical manifestations of IBD， IBD is closely related to the lung and the intestine. In terms of therapeutic principles， IBD can be treated by tonifying the spleen and replenishing the lung， which highlights the treatment of the intestine from the lung. To be specific， in time of tonifying the spleen and removing dampness， the intestine is regulated by tonifying the spleen and replenishing the lung. Shenling Baizhusan， a commonly used classical prescription for IBD， is mainly potent in replenishing Qi， invigorating the spleen， draining dampness， checking diarrhea， and especially "reinforcing earth to generate metal". It can enhance the function of the lung through "reinforcing earth to generate metal"， which in turn regulates the intestine and promote the improvement of IBD. The present study clarified the mechanism of Shenling Baizhusan in regulating the intestine by tonifying the spleen and replenishing the lung. On the basis of modern research， its therapeutic effect on IBD was achieved through multiple links， such as regulation of the level of inflammatory factors， immunoregulation， barrier function improvement via mucosal repair， and intestinal flora. The findings of this study are expected to provide new ideas for the regulation of the lung-spleen-large intestine axis in the syndrome differentiation and treatment of IBD and subsequent experimental research.

Exploration of IRES Elements within the ORF of the Coxsackievirus B3 Genome / 生物医学与环境科学（英文）

Objective@#This study aimed to identify internal ribosome entry sites (IRESs) in the open reading frame (ORF) of the Coxsackievirus B3 (CVB3) genome.@*Methods@#The sequences of P1, P2, or P3 of the CVB3 genome or the truncated sequences from each antithymocyte globulin (ATG) to the end of the P1, P2, or P3 gene were inserted into the pEGFP-N1 vector. After transfection, possible IRES-dependent green fluorescent protein (GFP)-fused proteins were detected by anti-GFP western blotting. The sequences of possible IRESs were inserted into specific Fluc/Rluc bicistronic vectors, in which the potential IRESs were determined according to the Fluc/Rluc activity ratio. Expression of Fluc and Rluc mRNA of the bicistronic vector was detected by RT-qPCR.@*Results@#After transfection of full length or truncated sequences of the P1, P2, or P3 plasmids, six GFP-fused protein bands in P1, six bands in P2 and nine bands in P3 were detected through western blotting. Two IRESs in VP2 (1461-1646 nt) and VP1 (2784-2983 nt) of P1; one IRES in 2C (4119-4564 nt) of P2; and two IRESs in 3C (5634-5834 nt) and 3D (6870-7087 nt) of P3 were identified according to Fluc/Rluc activity ratio. The cryptic promoter was also excluded by RT-qPCR.@*Conclusion@#Five IRESs are present in the CVB3 coding region.

Establishment of real-time fluorescent quantitative PCR for detection of torque teno virus types 7, 8 and 10 / 中华微生物学和免疫学杂志

Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.

Commelina communis L. extract inhibits hypoxia/reoxygenation-induced apoptosis of cardiomyocytes and expression of inflammatory factors through regulating NADPH oxidase 4 (Nox4) / 中华微生物学和免疫学杂志

Objective:To investigate the effects of Commelina communis L. on hypoxia/reoxygenation (H/R)-induced apoptosis of myocardial cells and expression of inflammatory factors through regulating NADPH oxidase 4 (Nox4). Methods:H9c2 cells were subjected to H/R to establish the injury model. These cells were then treated with different concentrations of Commelina communis L. extract. Cell activity and apoptosis were measured by MTT method and flow cytometry, respectively. Superoxide dismutase (SOD), malondialdehyde (MDA) and lactate dehydrofenase (LDH) levels were detected. ELISA was performed to detect TNF-α, IL-1β and IL-6 levels. Western blot was used to detect the expression of Bcl-2-associated X protein (Bax), cysteine-containing aspartic protease 3 (Caspase-3) and Nox4. Real-time quantitative PCR (qPCR) was used to measure the expression of Nox4 at mRNA level. H9c2 cells were transfected with si-Nox4 and subjected to H/R. Changes in the activity and apoptosis of H9c2 cells, and the expression of SOD, MDA, LDH and inflammatory factors were observed after inhibiting Nox4 expression. H9c2 cells transfected with pcDNA3.1-Nox4 were treated with Commelina communis L. extract and subjected to H/R to evaluate the effects on the cell activity and apoptosis, as well as the expression of SOD, MDA, LDH and inflammatory factors. Results:The survival rate and SOD activity of H9c2 cells exposed to H/R were significantly reduced, but the apoptosis rate, the levels of Caspase-3, Bax protein, MDA, LDH TNF-α, IL-1β and IL-6, and the expression of Nox4 at both mRNA and protein levels were dramatically increased ( P<0.05). As with inhibition of Nox4 expression, Commelina communis L. extract at the concentrations of 10 μg/ml and 20 μg/ml could obviously increase the survival rate and SOD activity of H9c2 cells after H/R exposure, and reduce the apoptosis rate and the expression of Caspase-3, Bax protein, MDA, LDH TNF-α, IL-1β and IL-6, as well as Nox4 expression at mRNA and protein levels ( P<0.05). Overexpression of Nox4 reversed the effects of Commelina communis L. extract on promoting cardiomyocyte cell activity and SOD activity, inhibiting cell apoptosis, and downregulating the expression of Caspase-3, Bax protein, MDA, LDH, TNF-α, IL-1β and IL-6 in the H/R injury model. Conclusions:Commelina communis L. could protect against H/R-induced myocardial cell injury, improve cell activity, and inhibit cell apoptosis and the expression of inflammatory factors through regulating Nox4 expression.

Acupuncture therapy with point selection based on syndrome differentiation along the meridians for functional dyspepsia: a randomized controlled trial / 针灸推拿医学（英文版）

Objective: To observe the difference in clinical efficacy between acupuncture with point selection based on syndrome differentiation along the meridians and acupuncture at non-meridian and non-acupoint points for functional dyspepsia (FD). Methods: A total of 74 FD patients were randomized into an observation group and a control group, with 37 cases in each group. Both groups received acupuncture treatment. Zusanli (ST 36) and Neiguan (PC 6) were selected in the observation group, with Taichong (LR 3) and Neiting (ST 44) added for excess syndrome, and Gongsun (SP 4) and Yinlingquan (SP 9) added for deficiency syndrome. Four non-meridian and non-acupoint points were selected in the control group. The treatments in both groups were performed once a day with a 2-day break after 5 consecutive treatments, which constituted one treatment course. A total of 4 courses were performed. The scores of Nepean dyspepsia index (NDI) and Leeds dyspepsia questionnaire (LDQ) were recorded before and after treatment, and during follow-up (8, 12, 16, 20 and 24 weeks after recruitment) to assess the clinical efficacy. Results: The NDI scores in the two groups after treatment and at each time point during follow-up were higher than those before treatment (all P<0.05), and the LDQ scores were lower than those before treatment (all P<0.05). The NDI scores after treatment and at each time point during follow-up in the observation group were higher than those in the control group (all P<0.01); the total LDQ score and scores of upper abdominal pain, postprandial satiety and upper abdominal burning sensation after treatment and at each time point during follow-up in the observation group were significantly lower than those in the control group (P<0.01 or P<0.05).. Conclusion: Acupuncture with point selection based on syndrome differentiation along the meridians has a better curative effect than acupuncture at non meridian and non-acupoint points in the treatment of FD.

Effect of moxibustion at sensitized-acupoints on quality of life in patients with chronic superficial gastritis / 针灸推拿医学（英文版）

Objective: To compare the efficacy difference between moxibustion at sensitized-acupoints and non-sensitized- acupoints using the same group of acupoints. Methods: A total of 139 patients with chronic superficial gastritis were divided into a sensitized acupoint group (102 cases) and a non-sensitized acupoint group (37 cases) based on whether acupoint sensitization occurred. The SPSS version 19.0 statistical software propensity score matching function was used to balance the baseline data between the groups. Finally, 29 pairs of matched patients were included, namely 29 cases in the sensitized acupoint group and 29 cases in the non-sensitized acupoint group. Both groups were treated with moxibustion therapy. The treatment lasted for 30 min per time, and was performed every other day for 8 weeks. Changes in the traditional Chinese medicine (TCM) symptom score and the short-form 36-item health survey (SF-36) score in both groups were observed before and after treatment, as well as the clinical efficacy. Results: The covariates of age, course of disease, TCM symptom score and SF-36 score in the two groups were balanced after matching (all P>0.05). After treatment, the total effective rate was 100.0％ in the sensitized acupoint group and 79.3％ in the non-sensitized acupoint group. The difference in the total effective rate between the two groups was statistically significant (P<0.01). After treatment and at the 4-week follow-up, the TCM symptom scores in the sensitized acupoint group were significantly lower than those in the non-sensitized acupoint group (all P<0.01); the SF-36 scores in the sensitized acupoint group were significantly higher than those in the non-sensitized acupoint group (all P<0.01). Conclusion: With the same group of acupoints, the sensitized acupoints have a better therapeutic effect and long-term efficacy than the non-sensitized acupoints in the treatment of chronic superficial gastritis.

Effect of moxibustion at Feishu (BL 13) on airway inflammation in asthma model rats / 针灸推拿医学（英文版）

Objective: To observe the effect of moxibustion at Feishu (BL 13) on related inflammatory cells and inflammatory factors in the bronchoalveolar lavage fluid of asthma model rats, and to explore the mechanism of moxibustion in treating asthma. Methods: A total of 48 Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, a moxibustion group and a medication group, with 12 rats in each group. Except for the normal group, the rats in the other three groups were subjected to ovalbumin sensitization to stimulate the asthma. At the same time, rats in the moxibustion group received moxibustion at bilateral Feishu (BL 13), and rats in the medication group received dexamethasone by intragastric administration. Rats in the normal and the model groups only received the same fixation and normal saline by intragastric administration. After the interventions, the inspiratory resistance, the expiratory resistance, and the pulmonary compliance were measured for rats in each group; the numbers of the inflammatory cells in the bronchoalveolar lavage fluid were counted; the levels of the involved inflammatory factors in bronchoalveolar lavage fluid were detected; the pathological morphologies of the lung tissues were observed under light microscope. Results: After modeling, compared with the normal group, the rats in the model group showed obvious asthma attack-like response, significantly increased inspiratory resistance and expiratory resistance (both P<0.01), and significantly reduced pulmonary compliance (P<0.01); thickened tracheal wall and the narrowed tracheal lumen observed under the light microscope; infiltration of inflammatory cells and increased eosinophils in and around the tracheal wall; significantly increased total number of inflammatory cells and proportion of eosinophils in the bronchoalveolar lavage fluid (all P<0.01); significantly reduced levels of interleukin (IL)-10, IL-12 and interferon (IFN)-γ (all P<0.01), and significantly increased levels of IL-4, IL-5 and tumor necrosis factor (TNF)-α (all P<0.01) in the bronchoalveolar lavage fluid. After intervention, compared with the model group, rats in the moxibustion and the medication groups showed significantly reduced asthma-like reaction, pathological morphological damage of lung tissue, inspiratory resistance and expiratory resistance (all P<0.01); significantly increased pulmonary compliance (P<0.01); significantly reduced total number of inflammatory cells, proportion of eosinophils, levels of IL-4, IL-5 and TNF-α in the bronchoalveolar lavage fluid (P<0.05 or P<0.01), while significantly increased IL-12 and IFN-γ levels (all P<0.01) in the bronchoalveolar lavage fluid; rats in the medication group also showed a significantly reduced IL-10 level in the bronchoalveolar lavage fluid (P<0.01); there was no statistically significant difference between the moxibustion and the medication groups (all P>0.05). Conclusion: Both moxibustion at Feishu (BL 13) and intragastric administration of dexamethasone can improve the asthma attack-like symptoms of ovalbumin-sensitized rats; regulating the inflammatory cell numbers and the inflammatory factor contents in the lung may be one mechanism of moxibustion in treating asthma.

Significance and operation mode of moxibustion intervention for the group under quarantine after close contact with COVID-19 / 中国针灸

On the base of the idea of traditional Chinese medicine as "disease prevention", the mode and the protocol of the moxibustion intervention for the group under quarantine after close contact with coronavirus disease 2019 (COVID-19) were explored. The group under quarantine after close contact with COVID-19 was taken as the subjects. By the non-contact physician-patient communication network platform co-developed by China Association of Acupuncture-Moxibustion, Hunan Provincial Association of Acupuncture-Moxibustion, Data Center of China Academy of Chinese Medical Sciences and Yuge Medicine Company, an exploratory randomized controlled trial was designed. A total of 100 cases were included and randomized into a moxibustion group and a conventional intervention group, 50 cases in each one. In the moxibustion group, moxibustion intervention was used. In the conventional intervention group, the conventional observation was adopted without moxibusiton intervention applied. The outcomes included the symptoms changes, e.g. anxiety, emotional disturbance, fatigue, headache and diarrhea, as well as whether quarantine release and the case confirmed or not, etc. The results were evaluated before intervention, in 14 days of intervention and 2 weeks after intervention separately. In this research, on the base of internet plus technology and with the internet communication platform adopted, through mobile phone WeChat App, it was to implement the subject screen, the random allocation and the instruction of moxibustion intervention as well as the quality control of patient's diary and data collection. It is anticipated that the significance and the implementation mode of moxibustion intervention can be assessed preliminarily for the group under quarantine after close contact with COVID-19.

A autophagy induced by non-stuctural protein 2B of human rhinovirus / 中华实验和临床病毒学杂志

Objective@#To study the intracellular location and autophagosome production of rhinovirus 16 2B protein using miniSOG labeling technique.@*Methods@#2B was fused with miniSOG and flag tags to construct pcDNA3.1-2B-miniSOG-flag plasmid, which was used to transfect HEK293 cells, LC3 protein was detected by western blot. The transfected cells were fixed, stained with DAB through the photooxidation activity of miniSOG, and used to prepare ultrathin sections. Localization of 2B-miniSOG protein in cells and ultrastructural changes of cells were observed under electron microscope.@*Results@#2B-miniSOG protein glows green under a fluorescence microscopy. Green flourescence coold be observed in the cells expressing 2B-miniSOG protein.LC-II protein increased in the cells transfected with pcDNA3.1-2B-miniSOG-flag. Under electron microscopy it was observed that 2B-miniSOG protein was located in the mitochondria, and a large number of vesicular structures appeared in the cytoplasm. Both autophagosomes and autophagic lysosomes can be observed.@*Conclusions@#Non-structural protein 2B of HRV16 can induce autophagy.

A preliminary study on thapsigargin promoting coxsackievirus B3 replication through activating transcription factor 6 pathway / 中华实验和临床病毒学杂志

Objective@#To investigate the effect of thapsigargin (TG) which can induce endoplasmic reticulum stress (ERS) on the replication of coxsackievirus B 3 (CV-B3).@*Methods@#After 10 MOI CV-B3 infected HeLa cells were exposed 0.25 μmol/L TG for 3 h, 6 h and 9 h, virus RNA of HeLa cells were extracted and viral replication was evaluated by real time PCR. After 0.25 μmol/L、0.08 μmol/L and 0.025 μmol/L TG exposed, the plaque of CV-B3 was used to confirm further replication of CV-B3. To verify TG induced ERS through three signal pathway, one of among PERK, ATF6 and IRE1 inhibitors GSK2656157, AEBSF and STF-083010, and 0.25 μmol/L TG were used in HeLa cells infected with 10 MOI CV-B3, replication of CV-B3 was evaluated by qRT-PCR.@*Results@#The stimulation of TG did not induce increase of virus replication after post-infection 3 h. However, TG induced replication of virus to increase 2.5 times after post-infection 6 h and 158.6 times after post-infection 9 h. And, the area of viral plaque was significantly increased. ATF6 inhibitors AEBSF significantly inhibited promotion of virus replication from TG.@*Conclusions@#TG can promote the replication of CV-B3 through ATF6 signal pathway.

Changes of endogenous small interfering RNA in H1-HeLa cells induced by human rhinovirus 16 infection / 中华实验和临床病毒学杂志

Objective@#To observe the changes of endogenous small interfering RNA (siRNA) in H1-Hela cells infected with human rhinovirus 16 (HRV 16).@*Methods@#To determine whether HRV16 infection induced the changes of siRNA, H1-HeLa cells were infected with HRV16 for 12 h, 24 h and 36 h, siRNAs were detected by high-throughput sequencing, second-generation sequencing) and qRT-PCR.@*Results@#The result showed that siRNA was generated differently at different time points post-infection, among which novel_sir907 and novel_sir1950 decreased at three time points. Further validation by qRT-PCR showed that novel_sir907 decreased at 12 h, 24 h and 36 h post-infection compared with the cell control, but novel_sir1950 increased at 12 h then decreased at 24 h and 36 h.@*Conclusions@#HRV16 infection induces changes endogenous siRNAs.

Multiple serotypes of Rhinovirus infection occurred in inpatients of an emergency department, Bengbu, Anhui province, in winter of 2018 / 中华实验和临床病毒学杂志

Objective@#To investigate the incidence of human rhinovirus (HRV) infection in hospitalized patients in emergency department.@*Methods@#A total of 94 emergency patients admitted to the emergency department from November to December of 2018 were enrolled in this study. The rhinovirus infection and related risk factors were analyzed.@*Results@#HRV infection occurred in 17 out of 94 hospitalized patients in emergency department, the infection rate was 18.09%; Multiple HRV serotypes were prevalent from November to December of 2018, which were A9, A10, A16, A31, A73, B42 and C3. Elderly patients are at high risk of rhinovirus infection; Rhinovirus infection may increase the course of inpatients.@*Conclusions@#From November to December of 2018, multiple serotypes of HRV infections in the emergency department of Bengbu, Anhui, and HRV easily infected elderly inpatients.

Establishment and verification of real-time fluorescent quantitative PCR detection system for ring virus 6 / 中华实验和临床病毒学杂志

Objective@#To establish a real-time quantitative PCR detection system for Torque teno virus (TTV) and verify the sensitivity and specificity of the detection system.@*Methods@#Primers and FAM-Eclipse probes were designed based on the TTV6 gene sequence registered in GenBank, and were to establish a real-time fluorescent quantitative PCR detecting way based on the FAM-Eclipse probe, the standard curve was constructed and sensitivity and specificity were analyzed.@*Results@#A quantitative PCR method for the specific detection of TTV6 were established that the standard curve equation was y=-3.0921x + 28.36, and the amplification efficiency and R2 were 99.6% and 1.000, respectively. The sensitivity of TTV6 was 1.0×10 copies/μl, and there was no cross-reactivity with other viruses. There was 1 case positive for TTV6 out of 56 throat swab samples from the patients with clinical respiratory infection.@*Conclusions@#The real-time fluorescent quantitative PCR for detecting TTV6 established by FAM-Eclipse probe had the advantages of high sensitivity and specificity. It provides an effective way for detection and quantification of viral content of TTV6 in clinical specimens.

Mechanism study of the transmission of moxibustion heat in human acupoint tissues / 针灸推拿医学（英文版）

Objective:To discuss the topical action characteristics of the biological transmission of moxibustion heat via temperature collection and numerical modeling.Methods:Temperature of moxibustion was measured at multiple points at a distance of 3 cm to obtain the moxibustion temperature field nephograms by the high-accuracy temperature measure array.Finite element analysis was used to imitate the three-dimensional dynamic distribution of temperature in acupoint tissues.Results:Through numerical analysis,the one-dimensional,two-dimensional and three-dimensional distributions of temperature in human acupoint tissues at 5 min of moxibustion were established.The result showed that moxibustion heat mainly transmitted from the surface of the tissue to the internal,and the influence of moxibustion heat decreased with the depth of the tissue.The analysis of the nephograms of acupoint tissue temperature at 5,10,15 and 20 min of moxibustion showed that with the increase of the moxibustion time,the temperature in acupoint tissues constantly rose,and the transmission depth of moxibustion heat also further expanded inside acupoint.Conclusion:By establishing the three-dimensional dynamic model of heat transmission inside acupoint tissues with the biological parameters of human tissues and the temperature values obtained,this study used finite element analysis software ANSYS 14.0 and discovered the rules in the transmission of heat in body tissues during moxibustion,and the features in moxibustion heat transmission (from the proximal to the distant) and heat penetration (from the surface to the internal).This study provides theoretical and experimental support for the application of moxibustion in clinical practice.