DDX21 mediates co-transcriptional RNA m6A modification to promote transcription termination and genome stability.

N6-methyladenosine (m6A) is a crucial RNA modification that regulates diverse biological processes in human cells, but its co-transcriptional deposition and functions remain poorly understood. Here, we identified the RNA helicase DDX21 with a previously unrecognized role in directing m6A modification on nascent RNA for co-transcriptional regulation. DDX21 interacts with METTL3 for co-recruitment to chromatin through its recognition of R-loops, which can be formed co-transcriptionally as nascent transcripts hybridize onto the template DNA strand. Moreover, DDX21's helicase activity is needed for METTL3-mediated m6A deposition onto nascent RNA following recruitment. At transcription termination regions, this nexus of actions promotes XRN2-mediated termination of RNAPII transcription. Disruption of any of these steps, including the loss of DDX21, METTL3, or their enzymatic activities, leads to defective termination that can induce DNA damage. Therefore, we propose that the R-loop-DDX21-METTL3 nexus forges the missing link for co-transcriptional modification of m6A, coordinating transcription termination and genome stability.

ssc-miR-204 regulates porcine preadipocyte differentiation and apoptosis by targeting TGFBR1 and TGFBR2.

MicroRNAs (miRNAs) take part in a variety of biological processes by regulating target genes. Transforming growth factor ß receptor 1 (TGFBR1) and TGFBR2 are crucial members of the TGF-ß family and are serine/threonine kinase receptors. The aim of this study was to explore the functions of ssc-miR-204 in porcine preadipocyte differentiation and apoptosis with regard to the TGFß/Smad pathway. We identified miRNAs predicted to target TGFBR1 and TGFBR2 using a database and selected ssc-miR-204 as a candidate miRNA. ssc-miR-204 overexpression dramatically reduced the levels of TGFBR1 and TGFBR2. However, after transfection with ssc-miR-204 inhibitor, TGFBR1 and TGFBR2 levels were dramatically increased. ssc-miR-204 overexpression dramatically promoted porcine preadipocyte differentiation and apoptosis. After transfection with ssc-miR-204 inhibitor, porcine preadipocyte differentiation and apoptosis were dramatically inhibited. After transfection with ssc-miR-204 mimics, Smad2, Smad3, Smad4, p-Smad2, and p-Smad3 protein levels significantly decreased, and adipogenesis was regulated by inhibiting the TGF-ß/Smad3 signaling pathway. Taken together, these results verified that ssc-miR-204 regulates porcine preadipocyte differentiation and apoptosis by targeting TGFBR1 and TGFBR2.

[Effects of Acupuncture Stimulation of Bilateral "Hegu" (LI 4) and "Taichong" (LR 3) on Learning-memory Ability, Hippocampal AP 42 Expression and Inflammatory Cytokines in Rats with Alzheimer's Disease].

OBJECTIVE: To observe the effect of acupuncture stimulation of bilateral "Hegu" (LI 4) and "Taichong" (LR 3, the so-called "Four Gate Points") on learning-memory ability, hippocampal interleukin-1 (IL-1) P and IL-2 and amyloid beta (Abeta) 42 levels in Alzheimer's disease (AD) rats,so as to reveal its underlying mechanism in improving AD. METHODS: Male SD rats were randomly divided into sham operation, model, medication and acupuncture groups (n = 12 rats in each group). The AD model was created by microinjection of streptozotocin (10 pL, 3 mg/kg) into the lateral ventricle (repeated the microinjection once two days later). Bilateral LR 3 and LI 4 were punctured with filiform needles and stimulated manually, once a day, 6 days a week for 4 weeks. The rats of the medication group were intragastric perfusion of Donepezil HOI (0.045 mg/kg), once a day for 4 weeks. The learning-memory ability was detected by Morris water maze swimming tests. The immunoactivity of hippocampal Abeta 42 was detected by immunohistochemistry, and the contents of IL-1 P and IL-2 in the hippocampus tissue were determined by ELISA. RESULTS: After modeling, the average escape latency of Morris water navigation task was significantly increased, and the target-platform crossing times of space probe trials were significantly reduced in the model group (P<0.05), suggesting a g-memory ability. After acupuncture intervention, the increased escape latency and the decreased target-platform crossing times were reversed, suggesting an improvement of the learning-memory. The hippocampal Abeta 42 immunoactivity and IL-1 beta content were significantly higher in the model group than in the sham operation group (P<0.05), but the hippocampal IL-2 content was markedly decreased in the model group (P<0. 05). Following the interventions, the increased Abeta 42 expression and IL-1 beta contents, and the decreased IL-2 contents in the hippocampus were also reversed in both the acupuncture and medication groups (P<0.05). CONCLUSION: Acupuncture may improve learning-memory ability in AD rats, which may be associated with its effects in reducing hippocampal Abeta13 42 expression and IL-1beta content and in up-regulating IL-2 level.

[Intervention of electroacupuncture for patients with impaired glucose tolerance].

OBJECTIVE: To explore the regulation on 2-hour postprandial blood glucose (2h PBG) for patients with impaired glucose tolerance (IGT) in the intervention with electroacupuncture. METHODS: Sixty cases of IGT were divided randomly into an electroacupuncture group and a blank control group, 30 cases in each one. In electroacupuncture group, electroacupuncture was applied to Shenshu (BL 23), Pishu (BL 20), Zusanli (ST 36) and Sanyinjiao (SP 6) in the intervention, lasting for 6 sessions. In blank control group, no any intervention was adopted. The levels of fasting blood-glucose (FBG), 2 h PBG with 75 g glucose and hemoglobin Alc (HbAlc) were observed before and after the intervention for the patients in electroacupuncture group, as well as in blank con trol group. RESULTS: The total effective rate was 76.7% (23/30) in electroacupuncture group, which was superior to that of 16.7% (5/30) in blank control group (P < 0.01). 2h PBG [(7.08 +/- 0.74) mmol/L] and HbAlc [(5.74 +/- 0.35)%] were down-regulated significantly after the intervention with electroacupuncture of 6 sessions in electroacupuncture group. The therapeutic effects of it were superior remarkably to those [(8.93 +/- 1.87) mmol/L, (5.97 +/- 0.591)%] in blank control group, indicating statistical significant difference (P < 0.01, P < 0.05). But, there was no any impact for the patients with normal FBG. CONCLUSION: Electroacupuncture may be the means to achieve the clinical effective intervention for the people with IGT and the approach in the prevention and treatment of diabetes at the early stage.

[Effects of different frequencies of electroacupuncture on blood glucose level in impaired glucose tolerance patients].

OBJECTIVE: To Investigate the clinical efficacy of electroacupuncture (EA) at different frequencies for patients with impaired glucose tolerance (IGT). METHODS: A total of 120 IGT outpatients were randomly divided into control, EA-5 Hz, EA-50 Hz, and EA-100 Hz groups (n = 30/group). EA (1 mA) was applied to bilateral Pishu (BL 20), Shenshu (BL23), Zusanli (ST 36) and Sanyinjiao (SP6) for 20 min, once daily for 60 sessions. Body mass index (BMI), fasting blood glucose (FBG) and 2-hour post-prandial blood glucose (2 h PBG) contents were detected by using BAYER Blood Sugar Analyzer and glycosylated haemoglobin (HbA1c) content was detected by enzymatic assay. RESULTS: Following the treatment, both HbA1c and 2 h PBG levels in the EA-5 Hz group were significantly lower than those of the control group and those of pre-treatment in the same one group (P < 0.05, P < 0.01). No significant differences were found between the EA-5 Hz and control groups, between pre-treatment and post-treatment in the EA-5 Hz group in BMI and FBG levels; between the EA-50 Hz and control groups, between the EA-100 Hz and control groups, and between pre-treatment and post-treatment in the EA-50 Hz and EA-100 Hz groups in BMI, FBG,2 h PBG and HbA1c levels (P > 0.05). CONCLUSION: Lower frequency EA of BL 20, BL 23, ST 36 and SP 9 can reduce HbA1c and 2 h PBG levels in IGT patients, suggesting a helpful effect of EA in controlling the development of diabetes.

[Clinical observation on acupuncture for treatment of reflux esophagitis of heat stagnation of liver and stomach type].

OBJECTIVE: To observe the clinical therapeutic effect of acupuncture for treatment of reflux esophagitis of heat stagnation of liver and stomach type. METHODS: Sixty-one cases were randomly divided into an acupuncture group (31 cases) and a medication group (30 cases). The acupuncture group was treated with needles at Zusanli (ST 36), Zhongwan (CV 12), Weishu (BL 21) and Neiguan (PC 6) mainly, once a day; and the medication group was treated with oral administration of 20 mg Omeprazole, once a day. The scores of clincial symptoms, comprehensive therapeutic effect, results of gastroscopy and pathology as well as recurrence rate etc. were observed before and after treatment. RESULTS: After treatment, the scores of symptoms significantly decreased in the two groups (both P < 0.01). The total effective rate of the acupuncture group was 90.3% (28/31), and 90.0% (27/30 )in the medication group, there was no statistical difference between two groups (P > 0.05); results of gastroscopy and esophageal mucosa pathology showed no statistical difference between two groups (both P > 0.05), the recurrence rate 12 weeks after treatment of 9.1% in the acupuncture group was lower than that of 42.9% in the medication grou p (P < 0.05). CONCLUSION: Acupuncture has preferable short and long-term therapeutic effects for treatment of reflux esophagitis of heat stagnation of liver and stomach type.

[Clinical study of the treatment of infant cerebral palsy with warm-reinforcing needling combined with rehabilitation training].

OBJECTIVE: To observe the therapeutic effect of warm-reinforcing needling combined with modern rehabilitation training for infant cerebral palsy (CP). METHODS: Forty cases of cerebral palsy children were randomly divided into rehabilitation (Rehab, n=19) and acupuncture (Acup) + Rehab (n=21) groups. Body acupoints used were Baihui (GV 20), Zusanli (ST 36), Quchi (LI 11), Huantiao (GB 30), Yinlingquan (GB 34), Sanyinjiao (SP 6), etc., and scalp-acupoints Zhisanzhen (the cross-point between the anterior hairline and the median line of the head, 3 cun bilateral to the crossing point), Naosanzhen [Nao-hu (GV 17), 1.5 cun bilateral to GV17], Balance Zone, Motor Zone, etc. . The treatment was conducted once every other day, with 3 months being a therapeutic course, 2 courses altogether. Rehabilitation training included physical training (PT), occupational therapy (OT) training, and speech training (ST), 5 times a week, with 3 months being a therapeutic course, 2 courses altogether. Gross motor function measure (GMFM) and comprehensive function (CF, including cognition, speech, motor, self-care and social-adaptable abilities) were evaluated. RESULTS: After the treatment, of the 19 and 21 cases in Rehab and Acup+ Rehab groups, 12 (63.16%) and 18 (85.71%) experienced marked improvement, 7 (36.84%) and 3 (14.29%) failed, with the effective rates being 63.16% and 85.71% respectively. The therapeutic effect of Acup+ Rehab group was markedly superior to that of Rehab group (P < 0.05). Self-comparison of two groups showed that the scores of CF and GMFM increased significantly in comparison with pre-treatment (P < 0.01), and the scores of F and GMFM of Acup + Rehab group were obviously higher than those of Rehab group (P < 0.01). CONCLUSION: Rehabilitation training combined with acupuncture can apparently improve CP children's motor and comprehensive functions, and the therapeutic effect of AcupWarm-reinf Rehab is evidently superior to that of simple rehabilitation training therapy.