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.

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*

The intestinal is the main and most important part of human flora settlement.The factors affecting the composition and diversity of intestinal flora in early life mainly include gestational age, delivery mode, feeding mode, antibiotic exposure, etc.The occurrence of neonatal late-onset sepsis(LOS)is associated with intestinal flora imbalance.The use of probiotics to regulate intestinal flora can reduce the incidence and mortality of LOS, which is helpful for treatment and prevention of LOS.

Objective:To analyze the potential risk factors of periventricular-intraventricular hemorrhage(PIVH)in premature infants.Methods:A retrospective study was conducted on clinical data of 279 premature infants admitted to the Affiliated Hospital of Guizhou Medical University From January 1, 2019 to December 31, 2019, who completed cranial ultrasound during hospitalization.According to the cranial ultrasound with or without PIVH, the cases were divided into PIVH group and non-PIVH group.The premature infants with PIVH were divided into severe PIVH(grade Ⅲ and Ⅳ)group and mild PIVH(grade Ⅰand Ⅱ)group according to the PIVH grades.A total of 25 factors, which may influnce PIVH, were analyzed by univariate analysis, and then multivariate Logistic stepwise regression analysis(stepwise backwards method)was performed to determine the major risk factors.Results:(1)A total of 279 premature infants were included in the study, 133 of them in PIVH group, and 146 of them in non-PIVH group.Univariate analysis showed that there were statistically significant differences in 14 factors between two groups, including full treatment of antenatal steroid, gestation age, birth weight, neonatal asphyxia, hypothermia, early onset sepsis, metabolic acidosis, hypernatremia, anemia, respiratory distress syndrome, noninvasive ventilation, invasive ventilation, invasive ventilation within 72 hours after birth, and lumbar puncture within 72 hours after birth( P<0.05). Multivariate analysis showed that gestational age( OR=0.709, 95% CI 0.602-0.835), and full treatment of antenatal steroid( OR=0.354, 95% CI 0.189-0.664) were protective factors for PIVH in premature infants, while neonatal asphyxia( OR=2.425, 95% CI 1.171-5.023), hypothermia( OR=2.097, 95% CI 1.088~4.041), early onset sepsis( OR=12.898, 95% CI 1.433-115.264), metabolic acidosis( OR=2.493, 95% CI 1.398-4.442), invasive ventilation within 72 hours after birth( OR=5.408, 95% CI 1.156-25.297), lumbar puncture within 72 hours after birth ( OR=5.035, 95% CI 1.269-19.993) were independent risk factors for PIVH in premature infants( P<0.05). (2) Among 133 cases of premature PIVH, 20 cases were severe PIVH and 13 cases were mild PIVH.Univariate analysis showed that there were statistically significant differences in 5 factors between two groups, including antenatal magnesium sulfate, gestation age, early onset sepsis, abnormal coagulation, and lumbar puncture within 72 hours after birth.Multivariate analysis showed that early onset sepsis( OR=4.392, 95% CI 1.343-14.367) and abnormal coagulation( OR=3.502, 95% CI 1.234-9.867) were independent risk factors for severe PIVH in premature infants( P<0.05). Conclusion:Gestational age is negatively correlated with the occurrence of PIVH in premature infants, and completion of more than a course of treatment for antenatal dexamethasone is an independent protective factor of PIVH in premature infants.Neonatal asphyxia, metabolic acidosis, hypothermia(<35 ℃), early onset sepsis, invasive ventilation within 72 hours after birth, and lumbar puncture within 72 hours after birth are independent risk factors for PIVH in premature infants.Abnormal coagulation and early onset sepsis are independent risk factors for severe PIVH in premature infants.

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.

Virus infection induces the production of type I interferons (IFNs). IFNs bind to their heterodimeric receptors to initiate downstream cascade of signaling, leading to the up-regulation of interferon-stimulated genes (ISGs). ISGs play very important roles in innate immunity through a variety of mechanisms. Although hundreds of ISGs have been identified, it is commonly recognized that more ISGs await to be discovered. The aim of this study was to identify new ISGs and to probe their roles in regulating virus-induced type I IFN production. We used consensus interferon (Con-IFN), an artificial alpha IFN that was shown to be more potent than naturally existing type I IFN, to treat three human immune cell lines, CEM, U937 and Daudi cells. Microarray analysis was employed to identify those genes whose expressions were up-regulated. Six hundred and seventeen genes were up-regulated more than 3-fold. Out of these 617 genes, 138 were not previously reported as ISGs and thus were further pursued. Validation of these 138 genes using quantitative reverse transcription PCR (qRT-PCR) confirmed 91 genes. We screened 89 genes for those involved in Sendai virus (SeV)-induced IFN-β promoter activation, and PIM1 was identified as one whose expression inhibited SeV-mediated IFN-β activation. We provide evidence indicating that PIM1 specifically inhibits RIG-I- and MDA5-mediated IFN-β signaling. Our results expand the ISG library and identify PIM1 as an ISG that participates in the regulation of virus-induced type I interferon production.
Cells, Cultured ; Gene Library ; Humans ; Interferon Type I ; metabolism ; Interferon-beta ; genetics ; metabolism ; Proto-Oncogene Proteins c-pim-1 ; genetics ; Up-Regulation

OBJECTIVETo evaluate the differences in the clinical therapeutic effects on spasmodic hemiplegia after stroke among the alliance therapy of scalp acupuncture, music therapy combined with rehabilitation, the simple rehabilitation therapy and the combination of music therapy and rehabilitation.

METHODSA total of 76 patients of post-stroke spasmodic hemiplegia were randomized into a rehabilitation group (25 cases), a combination group with music therapy and rehabilitation (25 cases) and an alliance therapy group with scalp acupuncture, music therapy and rehabilitation (26 cases). In the rehabilitation group, the routine rehabilitation therapy was applied, including the removal of various incentives that cause spasm, the correction of body position and the physical therapy. In the combination group, the music therapy was added on the basis of the treatment as the rehabilitation group. The music physician used the rhythmic auditory stimulation, the patterned sensory enhancement and the therapeutic instrumental music playing to set up the task in the treatment. In the alliance therapy group, scalp acupuncture was added on the basis of the treatment as the combination group. The anterior oblique line of vertex-tempora (MS 6) and the posterior oblique line of vertex-tempora (MS 7) on the contralateral side were selected and stimulated with penetrating needling technique. The needles were retained. During the needling retaining, the needles were rotated once every 10 min, for 2 min each time. The treatment was given one session a day, totally for 5 sessions a week, continuously for 4 weeks. The Fugl-Meyer assessment (FMA), Barthel index (BI) and the modified Ashworth scale (MAS) of the affected elbow and the passive knee movement at static condition were observed in the patients before and after treatment.

RESULTSThe results of FMA, BI and MAS were not different before treatment in the patients among the three groups (all>0.05), indicating the comparability among groups. After treatment, FMA and BI scores were all increased apparently in the three groups as compared with those before treatment (all<0.05). MAS grade was reduced remarkably as compared with that before treatment (all<0.05). After treatment, FMA and BI scores in the alliance therapy group were higher than those in the combination group and the rehabilitation group (all<0.05). FMA and BI scores in the combination group were higher than those in the rehabilitation group (both<0.05). MAS grade in the alliance therapy group was lower than those in the combination group and the rehabilitation group (both<0.05). MAS grade in the combination group was lower than that in the rehabilitation group (<0.05).

CONCLUSIONThe alliance therapy with scalp acupuncture, music therapy and rehabilitation achieve the remarkable clinical therapeutic effects on post-stroke spasmodic hemiplegia as compared with the routine rehabilitation and the combination of music therapy and rehabilitation.