OBJECTIVE: To observe the effects of Xingnao Kaiqiao acupuncture technique (for regaining consciousness and opening orifice) on methylation of N6-methyladenosine (m6A), and key methyltransferases and demethylases, so as to clarify the mechanism of acupuncture on cerebral ischemia-reperfusion injury (CIRI). METHODS: Of 68 male Sprague-Dawley rats of SPF grade, 15 rats were randomly selected as a sham-operation group, and the remaining rats were subjected to the model of middle cerebral artery occlusion using the suture ligation. CIRI was induced by ischemia for 2 h followed by reperfusion. Rats that failed to modeling or died were excluded. The rest 45 rats were randomly divided into three groups, i.e. model group, acupuncture group, and non-point acupuncture group, with 15 rats in each group. The rats in the acupuncture group were treated with acupuncture at bilateral "Neiguan" (PC6) and "Shuigou" (GV26). In the non-point acupuncture group, acupuncture was delivered at three non-points, located 3 mm below the bilateral midaxillary line and 3 mm lateral to the tip of the coccyx. One intervention was operated in these two acupuncture groups and the needles were retained for 30 min. Before modeling start and 2 h after ischemia, a laser speckle flowmeter was used to monitor the cerebral blood perfusion. In 2 h of ischemia and 24 h of reperfusion, the neurological behavioral score was evaluated. The volume of rat cerebral infarction was determined by triphenyltetrazolium chloride (TTC) staining, and the level of m6A methylation in ischemic cortical area was detected by Dot blot, and the protein and mRNA expression of the demethylase i.e. fat mass and obesity-associated protein (FTO), AlkB homolog 5 (ALKBH5) and key methyltransferases, i.e. methyltransferase-like 3 (METTL3), methyltransferase-like 14 (METTL14), and Wilms' tumor 1-associated protein (WTAP) in ischemic cortical area were detected by Western blot and real-time PCR. RESULTS: Cerebral blood perfusion decreased by>70% after 2 h ischemia. Compared with the sham-operation group, the neurobehavioral score and the percentage of cerebral infarction volume increased in the model group (P<0.01); the level of m6A methylation in the ischemic cortical area increased (P<0.01), the protein and mRNA expression of FTO decreased (P<0.01), and that of ALKBH5, METTL3, and METTL14 increased (P<0.01, P<0.05) in the model group. When compared with the model group and the non-point acupuncture group, the acupuncture group showed a decrease in the neurobehavioral score and the percentage of cerebral infarction volume (P<0.01), the level of m6A methylation in the ischemic cortical area was reduced (P<0.01, P<0.05), and the protein and mRNA expression of FTO was elevated (P<0.01). CONCLUSION Xingnao Kaiqiao acupuncture technique presents its protective effect on the brain in the rats with CIRI, which is related to up-regulating the expression of FTO and modulating m6A methylation.
Animals ; Rats, Sprague-Dawley ; Male ; Acupuncture Therapy ; Reperfusion Injury/genetics* ; Rats ; Brain Ischemia/genetics* ; Humans ; Adenosine/metabolism* ; Methylation ; Acupuncture Points ; Cerebral Cortex/metabolism*

Objective:To investigate impact of baicalein(BA)on airway inflammation in chronic obstructive pulmonary disease(COPD)rats by regulating TLR4/NF-κB pathway.Methods:Rats were randomly divided into model group(n=56)and sham operation group(n=10),COPD rat model was established by smoking and intratracheal instillation of LPS,successful modeling rats were randomly divided into COPD group,BA low dose(BA-L,100 mg/kg)group,BA high dose(BA-H,200 mg/kg)group,TLR4 activator LPS group(0.1 mg/kg)and BA-H+LPS group(0.1 mg/kg LPS+200 mg/kg BA).After 2 h,peak expiratory flow(PEF),forced expiratory volume in first second(FEV1),forced vital capacity(FVC)and other lung function indexes were measured;serum TNF-α,IL-6 and IL-8 levels were measured;bronchoalveolar lavage fluid(BALF)was collected to analyze number and classification of cells;lung tissues were isolated to analyze pathological changes and TLR4,p-NF-κB p65/NF-κB p65 expressions.Results:Com-pared with sham operation group,pathological damage in COPD group was serious,PEF,FEV1 and FVC were decreased significantly,serum IL-6,IL-8,TNF-α levels,leukocytes number,neutrophils and lymphocytes proportions,TLR4 and p-NF-κB p65/NF-κB p65 expressions were increased significantly(P<0.05);compared with COPD group,pathological damage in LPS group was further aggra-vated,PEF,FEV1 and FVC were reduced significantly,serum IL-6,IL-8,TNF-α levels,leukocytes number,neutrophils and lym-phocytes proportions,TLR4 and p-NF-κB p65/NF-κB p65 expressions were increased significantly(P<0.05),pathological damage in BA-L group and BA-H group gradually improved,PEF,FEV1 and FVC were increased significantly,serum IL-6,IL-8,TNF-α levels,leukocytes number,neutrophils and lymphocytes proportions,TLR4 and p-NF-κB p65/NF-κB p65 expressions were decreased significantly(P<0.05);LPS reversed protective effect of BA-H on COPD rats.Conclusion:BA can improve airway inflammation in COPD rats,which may be related to inhibition of TLR4/NF-κB pathway.

Objective:To investigate impact of baicalein(BA)on airway inflammation in chronic obstructive pulmonary disease(COPD)rats by regulating TLR4/NF-κB pathway.Methods:Rats were randomly divided into model group(n=56)and sham operation group(n=10),COPD rat model was established by smoking and intratracheal instillation of LPS,successful modeling rats were randomly divided into COPD group,BA low dose(BA-L,100 mg/kg)group,BA high dose(BA-H,200 mg/kg)group,TLR4 activator LPS group(0.1 mg/kg)and BA-H+LPS group(0.1 mg/kg LPS+200 mg/kg BA).After 2 h,peak expiratory flow(PEF),forced expiratory volume in first second(FEV1),forced vital capacity(FVC)and other lung function indexes were measured;serum TNF-α,IL-6 and IL-8 levels were measured;bronchoalveolar lavage fluid(BALF)was collected to analyze number and classification of cells;lung tissues were isolated to analyze pathological changes and TLR4,p-NF-κB p65/NF-κB p65 expressions.Results:Com-pared with sham operation group,pathological damage in COPD group was serious,PEF,FEV1 and FVC were decreased significantly,serum IL-6,IL-8,TNF-α levels,leukocytes number,neutrophils and lymphocytes proportions,TLR4 and p-NF-κB p65/NF-κB p65 expressions were increased significantly(P<0.05);compared with COPD group,pathological damage in LPS group was further aggra-vated,PEF,FEV1 and FVC were reduced significantly,serum IL-6,IL-8,TNF-α levels,leukocytes number,neutrophils and lym-phocytes proportions,TLR4 and p-NF-κB p65/NF-κB p65 expressions were increased significantly(P<0.05),pathological damage in BA-L group and BA-H group gradually improved,PEF,FEV1 and FVC were increased significantly,serum IL-6,IL-8,TNF-α levels,leukocytes number,neutrophils and lymphocytes proportions,TLR4 and p-NF-κB p65/NF-κB p65 expressions were decreased significantly(P<0.05);LPS reversed protective effect of BA-H on COPD rats.Conclusion:BA can improve airway inflammation in COPD rats,which may be related to inhibition of TLR4/NF-κB pathway.

Blinding is an important means to control and reduce measurement bias in clinical research， and blinding assessment is the main method to measure the success of the blinding method. By summarizing the current situation of blinding assessment in randomized controlled trials （RCT） of acupuncture， it was found that the report rate of blinding assessment by acupuncture RCT was relatively low， and the studies reporting blinding assessment had several problems， such as incomplete assessment individuals， unreasonable assessment questionnaires， and unscientific analysis methods， and the setting of the assessment time point is controversial. Given the above problems， this paper discussed the key elements of blinding assessment individuals， assessment questionnaires， assessment time points， and analysis methods. It is suggested that blinding assessment should be carried out on all blinded participants and personnel in the study； the assessment questionnaire should be designed by direct inquiry， with responses designed using three or more categorical options that include an "unclear" option； the early stage of the trial should be taken as the mandatory time point for assessment， integrating the evaluation index of the James blinding index and the Bang blinding index， in order to standardize the application of blinding assessment in acupuncture RCT and improve the quality of acupuncture clinical research.

Immune checkpoint inhibitors (ICIs) show unique advantages in the treatment of lung cancer, making the treatment of lung cancer enter the era of immunotherapy, but ICIs will also have adverse reactions, and the incidence of immune-induced hematological toxicity is not very high. Immunotherapy-induced thrombocytopenia is a rare adverse event.We report one case of thrombocytopenia induced by ICIs and review the literature on thrombocytopenia associated with ICIs and discuss the clinical features, possible mechanisms, and optimal treatment. 
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Humans ; Purpura, Thrombocytopenic, Idiopathic/drug therapy* ; Lung Neoplasms/drug therapy* ; Thrombocytopenia/chemically induced* ; Antibodies, Monoclonal, Humanized/adverse effects*

Objective:To investigate the effect of astragaloside Ⅳ on the apoptosis of thyroid cells in Hashimoto's thyroiditis(HT)rats and Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil containing kinase 2(ROCK2)pathway.Methods:The HT rat model was induced by subcutaneous injection of thyroglobulin combined with high iodine drinking water and randomly divided into model group,astragaloside(80 mg/kg)group,Rhosin(RhoA inhibitor,40 mg/kg)group,astragaloside Ⅳ(80 mg/kg)+ Rhosin(40 mg/kg)group(12 rats in each group),another 12 SD rats were selected and drank water normally and injected the same dose of saline subcutaneously as control group.After the drugs were grouped and processed,the serum anti-thyroglobulin antibody(TGAb),anti-thyroid peroxidase antibody(TPOAb)levels and the inflammatory factors IL-6,IL-17,IL-1β contents were measured by ELISA kits;the pathological changes of thyroid tissue in each group were detected by hematoxylin-eosin(HE)staining;the apopto-sis rate of rat thyroid cells in each group were detected by TUNEL staining;the expressions of RhoA/ROCK2 pathway proteins in thy-roid tissues of rats in each group were detected by Western blot.Results:Compared with the control group,the thyroid follicles in the model group had abnormal structure,some atrophy or disappearance,disordered arrangement,surrounding inflammatory cell infiltra-tion,and obvious pathological damage to the thyroid tissue,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,and thyroid tissue RhoA and ROCK2 protein expression levels were significantly increased(P<0.05);compared with model group,the pathological damage of the thyroid tissue of rats in the drug intervention group were reduced,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,and thyroid tissue RhoA and ROCK2 protein expression levels were decreased(P<0.05);compared with astragaloside Ⅳ group and the Rhosin group respectively,the pathological damage of the thyroid tissue of rats in the astragaloside Ⅳ+Rhosin group were further reduced,the serum TGAb,TPOAb,IL-6,IL-17 and IL-1β levels,thyroid cell apoptosis rate,thyroid tissue RhoA and ROCK2 protein expression levels were decreased(P<0.05).Conclusion:Astragaloside Ⅳ may down-regulate the expression of RhoA/ROCK2 pathway to reduce the inflammatory injury of thyroid tissue,inhib-it thyroid cell apoptosis,and improve the symptoms of HT in rats.

To investigate the effects of ()infection on autonomic nervous function and calcitonin gene-related peptide in patients with functional dyspepsia(FD). Thirty-one patients with FD matching Rome Ⅳ criteria were included and divided into -positive group and -negative group.All patients were evaluated by Symptom Index of Dyspepsia(SID),Nepean Dyspepsia Index(NDI),and Hospital Anxiety and Depression Scale(HADS).Their heart rate variability(HRV)and calcitonin gene-related peptide(CGRP)level were also measured. There were no significant differences in SID(=-0.858, =0.858),NDI(=-1.464, =0.143),and Hospital Depression Scale score(=0.699, =0.485).However,the Hospital Anxiety Scale score was significantly higher in -positive group than the -negative group(=-2.470, =0.014).The level of CGRP in -positive group[(0.999±0.274)ng/ml]was significantly higher than that in the -negative group[(0.812±0.172)ng/ml;=2.238, =0.033].HRV data showed no significant difference between these two groups at very low frequency(=-1.210, =0.236),low frequency(LF)(=0.419, =0.678),high frequency(HF)(=0.612, =0.546),LF/HF(=-0.882, =0.399),and total power(=-0.963, =0.344). In FD patients,patients with -positive FD patients have higher depression and CGRP levels than those without infection,although their dyspepsia symptoms and HRV show no notable changes.
Anxiety ; Calcitonin ; Calcitonin Gene-Related Peptide ; Dyspepsia ; Helicobacter Infections ; Helicobacter pylori ; Humans

Objective: To explore the changes and clinical significance of serum prolactin and estrogen levels in women with autoimmune thyroid diseases. Methods: From January 2018 to December 2018, 76 newly diagnosed female patients with autoimmune thyroid diseasein outpatient and inpatient clinics of the People's Hospital of Hebei Province were selected as study group, including 40 cases of Graves' disease and 36 cases of hashimoto's thyroiditis.And 60 healthy women with age matched were selected as control group.Serum estrogen, prolactin, thyroid hormone and their antibodies, IL-2, IL-6, IL-10 and other related indicators were determined before and after treatment, and the correlation analysis was performed. Results: The levels of estrogen[(302.85±78.62)ng/L], prolactin [(15.98±4.18)μg/L], IL-2 [(224.45±61.28)ng/L], IL-6 [(211.46±67.25)ng/L] in the study group were all higher than those in the control group [(228.4±71.38)ng/L, (10.35±3.21)μg/L, (120.34±38.27)ng/L, (165.51±50.09)ng/L], and the IL-10 level in the study group was lower than that in the control group [study group: (15.65±4.86)ng/L; control group: (20.12±4.83)ng/L] , there were statistically significant difference between the two groups(t=5.78, 3.11, 8.98, 3.94, all P<0.05). After treatment, the levels of estrogen [(237.11±70.42)ng/L], prolactin[(10.40±3.32)μg/L], IL-2[(122.67±38.99)ng/L], IL-6[(166.24±52.09)ng/L] in the study group were significantly decreased compared with before treatment(t=5.56, 3.76, 9.04, 4.02, all P<0.05), and the IL-10 level [(19.96±4.93)ng/L] was significantly increased (t=3.95, P<0.05). However, there was no statistically significant difference in thyroid antibody between the two groups (P>0.05). Estrogen level was positively correlated with prolactin, IL-2 and IL-6(r=0.327, 0.298, 0.336, all P<0.01), and negatively correlated with IL-10(r=-0.285, P<0.05). Conclusion Elevated levels of estrogen and prolactin can stimulate the production and release of inflammatory cytokines, thus affecting the level of thyroid hormone, which is closely related to the female autoimmune thyroid disease.

Objective To investigate the clinicopathological features and correlation between synchronous multiple early gastric cancer (SMEGC)and single early gastric cancer (EGC).Methods From January 2008 to December 2016,the clinical data of 994 patients with EGC who underwent open or laparoscopic gastrectomy surgery were collected from the electronic medical data base of the First Affiliated Hospital of Nanjing Medical University and Xuzhou No.1 People's Hospital.The data of patients including gender,age,tumor morphologys,tumor location,tumor size,histological type,depth of invasion,lymph nodes metastasis,lymphovascular metastasis,peripheral nerve invasion,and blood types were analyzed.T test and Chi square test were used for statistical analysis.Results Among 994 EGC patients,27 cases (2.7％) were SMEGC,and 967 cases (97.3％) were single EGC.The percentage of male and female of single EGC were 71.4％ (690/967) and 28.6％ (277/967),respectively;the percentage of male and female of SMEGC were 88.9％ (24/27) and 11.1％ (3/27),respectively,and there was statistically significant difference in the gender composition ratio between single EGC and SMEGC (x2 =3.975,P=0.046).The incidence of ulcer in single EGC and SMEGC were 50.6％ (489/ 967) and 29.6 ％ (8/27),respectively,and the difference in the incidence of ulcers between single EGC and SMEGC was statistically significant (x2 =4.653,P=0.031).There were no statistically significant differences between single EGC and SMEGC in gross morphology,depth of invasion,lymph nodes metastasis,lymphovascular metastasis,peripheral nerve invasion,tumor location,pathological type and blood types (all P＞0.05).In the SMEGC patients,the incidence of main lesions invading the mucosa was 48.1％ (13/27) and submucosa invasion was 51.9％ (14/27);and for minor lesions,the corresponding incidences were 77.8％ (21/27) and 22.2％ (6/27),respectively,and the difference was statistically significant (x2 =5.063,P＜0.05).There were no statistically significant differences between the main lesions and minor lesions in tumor size,pathological type,with or without ulcers,gross morphology and tumor location (all P＞0.05).Conclusions The main risk factors of SMEGC are male and no ulcerative lesions.The clinicopathological features are similar between main lesions and minor lesions in SMEGC.

OBJECTIVETo discuss the effects on detrusor hyperreflexia treated with ginger-salt-isolated moxibustion at "Shenque" (CV 8) and its mechanism.

METHODSThirty female adult SD rats were selected. The model of detrusor hyperreflexia was prepared with complete spinal transection at T, of which, 20 rats were randomized into a model group (10 rats) and a moxibustion group (10 rats). A sham-operation group (10 rats) was set up for sham-spinal transection. In the moxibustion group, when urine incontinence occurred (about in 2 weeks of modeling), the ginger-salt-isolated moxibustion at "Shenque" (CV 8) was given, 3 moxa cones each time, once a day, continuously for 7 days. After treatment, in each group, the urodynamic parameters were determined, after which, the bladder detrusor was collected. Western blot was used to determine the protein expressions of M2 and M3 receptors.

RESULTSCompared with the sham-operation group, the micturition interval was shortened apparently (<0.01); the maximal bladder pressure was increased apparently (<0.01); the protein expression of M2 receptor in the detrusor was increased significantly (<0.05) and that of M3 receptor had no apparent change (>0.05) in the rats of the model group. Compared with the model group, the micturition interval was longer apparently (<0.01), the maximal bladder pressure was reduced apparently (<0.01), the protein expression of M2 receptor in the detrusor was reduced significantly (<0.05) and that of M3 receptor had no apparent change (>0.05) in the rats of the moxibustion group.Compared with the sham-operation group, the results of the above indicators were not different significantly in the moxibustion group (all>0.05).

CONCLUSIONSThe ginger-salt-isolated moxibustion at "Shenque" (CV 8) suppresses the overactive bladder in the rat with spinal transection and its effect mechanism is possibly relevant with reducing the protein expression of detrusor M2 and inhibiting the excessive contraction of the detrusor.