Novel roles of an intragenic G-quadruplex in controlling microRNA expression and cardiac function.

Simultaneous dysregulation of multiple microRNAs (miRs) affects various pathological pathways related to cardiac failure. In addition to being potential cardiac disease-specific markers, miR-23b/27b/24-1 were reported to be responsible for conferring cardiac pathophysiological processes. In this study, we identified a conserved guanine-rich RNA motif within the miR-23b/27b/24-1 cluster that can form an RNA G-quadruplex (rG4) in vitro and in cells. Disruption of this intragenic rG4 significantly increased the production of all three miRs. Conversely, a G4-binding ligand tetrandrine (TET) stabilized the rG4 and suppressed miRs production in human and rodent cardiomyocytes. Our further study showed that the rG4 prevented Drosha-DGCR8 binding and processing of the pri-miR, suppressing the biogenesis of all three miRs. Moreover, CRISPR/Cas9-mediated G4 deletion in the rat genome aberrantly elevated all three miRs in the heart in vivo, leading to cardiac contractile dysfunction. Importantly, loss of the G4 resulted in reduced targets for the aforementioned miRs critical for normal heart function and defects in the L-type Ca2+ channel-ryanodine receptor (LCC-RyR) coupling in cardiomyocytes. Our results reveal a novel mechanism for G4-dependent regulation of miR biogenesis, which is essential for maintaining normal heart function.

Homocysteine as a potential predictive factor for high major adverse cardiovascular events risk in female patients with premature acute coronary syndrome.

We aimed to investigate the correlation of homocysteine (Hcy) level with clinical characteristics, and explore its predictive value for major adverse cardiovascular events (MACE) risk in female patients with premature acute coronary syndrome (ACS).The serum Hcy level was detected from 1299 female patients with premature ACS. According to the tertile of Hcy level, patients were divided into 3 groups: lowest tertile group (≤9.1 µmol/L), middle tertile group (9.2-11.6 µmol/L) and highest tertile group (>11.6 µmol/L). MACE incidence was recorded and MACE-free survival was caculated with the median follow-up duration of 28.3 months.Increased Hcy correlated with older age (P < .001), higher creatinine level (P < .001), and enhanced uric acid level (P = .001), while reduced fasting glucose concentration (P < .001). MACE incidence was 10.7% and it was highest in highest tertile group (22.1%), followed by middle tertile group (7.7%) and lowest tertile group (2.4%) (P < .001). Receiver operating characteristic curve showed that Hcy distinguished MACE patients from non-MACE patients with the area under the curve of 0.789 (95% CI: 0.742-0.835). Kaplan-Meier curves revealed that MACE-free survival was shortest in Hcy highest tertile group, followed by middle tertile group and lowest tertile group (P < .001). Multivariate Cox analyses further showed that higher Hcy level was an independent predictive factor for poor MACE-free survival (middle tertile vs lowest tertile (P = .001, HR: 3.615, 95% CI: 1.661-7.864); highest tertile vs lowest tertile (P < .001, HR: 11.023, 95% CI: 5.356-22.684)).Hcy serves as a potential predictive factor for increased MACE risk in female patients with premature ACS.

Clinical observation on treatment of clearing the Governor Vessel and refreshing the mind needling in neural development and remediation of children with cerebral palsy.

OBJECTIVE: To investigate the effects of clearing the Governor Vessel and refreshing the mind needling in neural development and remediation of children with cerebral palsy. METHODS: A total of 200 cases of children with cerebral palsy were randomly assigned to the treatment group (100 patients) and the control group (100 patients). The treatment group was given the combined therapy of acupuncture and rehabilitation training, and the chosen acupoints were 13 points of the Governor Vessel, Shenshu (BL 23), Taixi (KI 3), Yanglingquan (GB 34), Zusanli (ST 36) and Sanyinjiao (SP 6), and points of refreshing the mind were also selected, which included puncturing Shenting (GV 24) toward Qianding (GV 21), puncturing Qianding (GV 21) toward Baihui (GV 20), puncturing Baihui (GV 20) toward Naohu (GV 17) and Sishencong (Ex-HN 1). The control group was only treated with rehabilitation training. A contrastive analysis of the therapeutic effect between acupuncture combined with rehabilitation training and rehabilitation training alone was made after a treatment course of 3 months. The Gross Motor Function Measure (GMFM) and Beijing Gesell Developmental Scale were adopted to assess the neural development and rehabilitation outcomes of the two groups. In addition, skull CT/MRI was adopted to evaluate the plerosis of injured cerebral nerve after treatment. RESULTS: The total effective rate in treatment group was 87% (87/100), significantly higher than the 55% (55/100) in the control group. The children's development quotient (DQ) tested by Gesell Developmental Scale and scores tested by GMFM in the treatment group were obviously higher than those in the control group (P<0.01). The improving and curing rates presented by skull CT/MRI in the treatment group were higher than those in the control group (P<0.01). CONCLUSIONS: Clearing the Governor Vessel and refreshing the mind needling could accelerate the recovery of injured brain nerve and the reconstruction of brain function. The acupuncture therapy could ameliorate both the motor development and cognitive development. On the other hand, the forward curative effect of acupuncture combined with rehabilitation training was significantly better than the rehabilitation training alone.

Up-regulation of PIK3CA promotes metastasis in gastric carcinoma.

AIM: To explore expressions of PIK3CA in the progression of gastric cancer from primary to metastasis and its effects on activation of phosphatidylinositol 3-kinase (PI3K)/Akt pathway. METHODS: mRNA and protein levels of PIK3CA were assessed, respectively, by real-time quantitative polymerase chain reaction and immunohistochemistry in specimens of normal gastric mucosa, primary foci and lymph node and distant metastasis of gastric cancer. Akt and phosphorylated Akt protein were also examined by Western blotting in these tissues, in order to analyze the effect of PIK3CA expression level changes on the activation of PI3K/Akt signaling pathway. RESULTS: PIK3CA mRNA in lymph node metastasis were approximately 5 and 2 folds higher, respectively, than that in the corresponding normal gastric mucosa and primary gastric cancer tissues (P < 0.05), while no statistical significance was found compared with distant metastasis. Immunohistochemically, PIK3CA protein expression was discovered in 7 (35%) specimens of 20 primary foci vs 10 (67%) of 15 of lymph node metastasis or 11 (61%) of 18 of distant metastasis (35% vs 67%, P = 0.015; 35% vs 61%, P = 0.044). With the increased level of PIK3CA expression, the total Akt protein expression remained almost unchanged, but p-Akt protein was upregulated markedly. CONCLUSION: Increased expression of PIK3CA is expected to be a promising indicator of metastasis in gastric cancer. Up-regulation of PIK3CA may promote the metastasis of gastric cancer through aberrant activation of PI3K/Akt signaling.

[Studies on IFN-gamma-mediated reversion of CIK killing sensitivity to edited human lung cancer A549 cells].

AIM: To explore IFN-gamma-mediated reversion of CIK killing sensitivity to immunoedited lung cancer A549 cells. METHODS: RT-PCR and MTT methods were used to detect the effect on MICA mRNA expression induced by IFN-gamma in the edited A549 cells and the change of CIK killing sensitivity to A549 cells, respectively. RESULTS: Low expression of cell surface MICA and low killing sensitivity of CIKs were observed in edited A549 cells. IFN-gamma could significantly increase MICA mRNA expression in the edited A549 cells and improve CIK cell cytotoxicity. CONCLUSION: IFN-gamma could reverse CIK killing sensitivity to the edited A549 cells by enhancing the MICA mRNA expression.

[Effect of acupuncture-moxibustion combined with nerve growth factor on compensation of cerebral function in the children of cerebral palsy].

OBJECTIVE: To investigate the compensation of cerebral function in acupuncture for rehabilitation of cerebral palsy. METHODS: One hundred children of cerebral palsy were randomly divided into a treatment group and a control group, 50 cases in each group. The treatment group were treated with scalp acupuncture at the Motor Area, Foot Motor Sensory Area and Equilibrium Area, body acupuncture at Binao (LI 14), Fengchi (GB 20), Huantiao (GB 30), etc. and injection of nerve growth factor into Zusanli (ST 36), in combination with rehabilitation training; the control group were treated only with rehabilitation training. Their clinical therapeutic effects and recoveries of brain lesion detected by CT, SEPCT were investigated. RESULTS: The total effective rate was 84.0% in the treatment group better than 52.0% of the control group. After treatment, the development quotient (DQ) in the treatment group was higher than the control group (P<0.01). CONCLUSION: Acupuncture can promote compensation of cerebral function in the children of cerebral palsy.

[Effect of Fuzheng Peiben therapy on dendritic cell in patients with breast cancer].

OBJECTIVE: To investigate the repairing mechanism of Fuzheng Peiben Therapy on cellular immunofunction in patients with breast cancer on cellar and molecular level. METHODS: The tumor tissue and axillary lymph node samples were evaluated with the flow cytometry for CD83, CD80, CD86 before and after neoadujvant therapy. RESULTS: The levels of CD83, CD80, CD86 were singificantly decreased in group A (treated with surgery only), B (treated with neoadujvant chemotherapy plus surgery) and C (treated with neoadujvant chemotherapy plus surgery with shenqi Fuzheng Injection) compared with that in normal team (P < 0.05 or 0.01); The levels of CD83, CD80, CD86 were significantly decreased in group A, B and C compared between before and after neoadujvant chemotherapy (P < 0.05 or 0.01); They were not significantly decreased between group A and group C (P > 0.05). CONCLUSION: The cellular immunity was inpaired considerably by neoadujvant chemotherapy. Neoadjuvant chemotherapy combined with Shenqi Fuzheng Injection can enhance the cellular immunity considerably.

Clinical study on treatment of mammary cancer by shenqi fuzheng injection in cooperation with chemotherapy.

OBJECTIVE: To study the effect of Shenqi Fuzheng Injection (SFI) on cellular immune in patients with mammary cancer (MC) after chemotherapy. METHODS: One hundred and ten patients with MC were randomly assigned to two groups. The 58 patients in the tested group were treated with SFI in cooperation with chemotherapy of CAF protocol (Cyclophosphamide, Doxorubicin and Fluorouracil), while the 52 patients in the control group were treated with chemotherapy of the same protocol alone. Changes of the patients' quality of life (QOF), adverse reaction that occurred, peripheral lymphocyte count and killing activity of single karyocyte before and after treatment between the two groups were compared. RESULTS: Patients' QOF elevating rate after treatment in the tested group and the control group was 34.5% and 13.5% respectively; The lowering of peripheral blood cell count of WBC, platelet and lymphocyte as well as that of the killing activity of single peripheral karyocyte on various kinds of MC cells were all milder and recovery sooner than those in the control group. CONCLUSION: SFI in combination with chemotherapy in treating MC could reduce the occurrence of adverse reaction to chemotherapy, improve clinical symptoms, elevate QOF and enhance immunity in patients with MC.