[Effect of LINC00174 on the Malignant Proliferation of Multiple Myeloma Cells and Its Related Mechanism].

OBJECTIVE: To explore the biological function of LINC00174 in multiple myeloma (MM). METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of LINC00174 and miR-150 in peripheral blood of MM patients and MM cell lines. EdU staining and flow cytometry were used to detect the effects of LINC00174 and miR-150 on the proliferation and apoptosis of MM cells. Western blot was used to detect the expressions of proliferation marker nuclear-related antigen Ki67, apoptosis-related protein cleaved caspase-3 and transcription factor forkhead box protein P1 (FOXP1). Bioinformatics and dual-luciferase reporter assay were used to verify the targeting relationship between LINC00174 and miR-150 and the targeting relationship between miR-150 and FOXP1. RESULTS: The level of LINC00174 was significantly increased in peripheral blood of MM patients and MM cell lines (P <0.05). Compared with NC-siRNA group, the expression of LINC00174 was significantly reduced in LINC00174-siRNA group, the proliferation of U266 cells was reduced, the apoptosis rate was significantly increased, the level of Ki67 protein was reduced, and the level of cleaved caspase-3 protein was increased (all P <0.05). LINC00174 targeted regulation of the expression of miR-150. Compared with LINC00174-siRNA+NC inhibitor group, the expression of miR-150 in U266 cells in LINC00174-siRNA+miR-150 inhibitor group was significantly reduced, the cell proliferation was enhanced, the apoptosis rate was reduced, the level of Ki67 protein was increased, and the level of cleaved caspase-3 was decreased (all P <0.05). FOXP1 is the target gene of miR-150. Compared with NC mimic group, the expression of FOXP1 protein in miR-150 mimic group was significantly reduced, the cell proliferation was reduced, the apoptosis rate was significantly increased, Ki67 protein level was decreased, and the level of cleaved caspase-3 was increased. Compared with miR-150 mimic + vector group, the expression of FOXP1 protein in miR-150 mimic + pcDNA-FOXP1 group was significantly increased, the cell proliferation was enhanced, the apoptosis rate was reduced, the level of Ki67 protein was increased, and the level of cleaved caspase-3 was decreased (all P <0.05). CONCLUSION: LINC00174 promotes the proliferation of MM cells and inhibits cell apoptosis by regulating the miR-150/ FOXP1 axis.

A new technique for performing interstitial implants for gynecologic malignancies using transvaginal ultrasound guidance.

Objectives: This study concerns a new technique that aims to achieve precise interstitial brachytherapy of pelvic recurrent tumors under transvaginal ultrasound (US) guidance, enhance the conformity index of the brachytherapy (BT), and improve the curative effect of radiotherapy for gynecological oncology patients with pelvic relapse. Methods: A real-time transvaginal US-guided interstitial implant device was developed to assist in implant BT. Prior to implant brachytherapy, the size and location of the tumor in the pelvis and the interrelationship with adjacent organs were first assessed with intracavitary ultrasound. The transvaginal US-guided interstitial implant device was then placed on the endoluminal ultrasound probe, the probe was oriented intravaginally to determine a safe needle path, the implant needle was placed into the needle passage of the device, and the implant needle was inserted into the tumor tissue in the direction guided by the ultrasound puncture guide line. After the implant needle was placed in place, the cover of the transvaginal US-guided interstitial implant device was opened perpendicular to the ultrasound probe, and the needle was separated from the ultrasound probe smoothly, and then the cover was re-covered for subsequent implantation. Results: In this study, 56 patients who underwent real-time transvaginal ultrasound-guided implantation for gynecologic oncology were enrolled, and insertion of 736 implant needles was completed. Among them, 13 patients had recurrent pelvic tumors after cervical cancer surgery and 6 patients had recurrent pelvic tumors after endometrial cancer surgery. Thirty-two patients who underwent radical radiation therapy for cervical cancer did not have adequate regression of parametrial invaded tissue after completion of standard EBRT treatment; and 5 patients had recurrent tumors in the radiation field after previous standard course of pelvic radiotherapy. The accuracy of the implant therapy was improved. The radiotherapy dose for recurrent pelvic masses was successfully increased, and the cumulative dose of external irradiation combined with BT was augmented to 80-100 Gy. The use of a new device for transvaginal implant for recurrent masses located in the lateral wall of the pelvic cavity was successful. Conclusion: This intravascular US-guided interstitial implant device can realize interstitial implant with the shortest path under transvaginal US guidance. With convenient operation, high precision, and good security, the device not only improves the accuracy of implant therapy, but it also reduces the risks of anesthesia and organ injury, so it is suitable for widespread promotion and use.

Quercitrin restrains the growth and invasion of lung adenocarcinoma cells by regulating gap junction protein beta 2.

Lung adenocarcinoma (LUAD) is the most prevalent subtype of non-small cell lung cancer (NSCLC) with high lethality, and quercitrin exhibits anticancer characteristics. Here, we attempted to uncover the anticancer activity of quercitrin in LUAD. In this work, quercitrin prohibited the cell viability and clone-formation of LUAD cells in vitro. Meanwhile, quercitrin treatment reduced the aggressive phenotypes in LUAD cells. Further, Gap Junction Protein Beta 2 (GJB2) expression was aberrantly higher in LUAD when compared within control tissue. The higher expression of GJB2 is associated with an inferior overall survival for patients with LUAD. Finally, the reintroduction of GJB2 offset the inhibiting influence of quercitrin in LUAD cells. Altogether, these findings disclosed that quercitrin suppressed the growth and metastatic-related traits of LUAD cells partly via regulating GJB2 expression.

[Effect of row-like puncture at sternocleidomastoid on peripheral facial palsy at recovery stage].

OBJECTIVE: To observe the effect of conventional acupuncture combined with row-like puncture at sternocleidomastoid on peripheral facial palsy at recovery stage. METHODS: A total of 60 patients with peripheral facial palsy at recovery stage were randomized into an observation group and a control group, 30 cases in each one. Acupuncture was applied at affected Cuanzhu (BL 2), Yangbai (GB 14), Sibai (ST 2), Quanliao (SI 18), Jiache (ST 6), Dicang (ST 4), Hegu (LI 4), Taichong (LR 3) and Zusanli (ST 36) in the control group. On the basis of the treatment in the control group, row-like puncture was applied at sternocleidomastoid (1 needle was punctured at muscle origin and insertion respectively, 3 to 4 needles were row-like punctured at the connection line of muscle origin and insertion). The treatment was given once a day, 5 times were as one course, with 2-day interval, totally 4 courses were required in the both groups. The house-brackmann (H-B) facial nerve function grade, facial nerve function rating system-dynamic view rating scale score and facial disability index (FDI) scale score [including scores of FDI physical function (FDIp) and FDI social life function (FDIs)] before and after treatment were observed, and the clinical efficacy was evaluated in the two groups. RESULTS: After treatment, the H-B facial nerve function grades were improved compared before treatment in the both groups (P<0.05), and that in the observation group was superior to the control group (P<0.05). After treatment, the scores of dynamic view rating scale and FDIp were increased (P<0.05), FDIs scores were decreased compared before treatment in the both groups (P<0.05); compared with the control group, the scores of dynamic view rating scale and FDIp were increased (P<0.05), FDIs score was decreased in the observation group (P<0.05). The total effective rate was 93.3% (28/30) in the observation group, which was superior to 73.3% (22/30) in the control group (P<0.05). CONCLUSION: Compared with conventional acupuncture, combination therapy with row-like puncture at sternocleidomastoid can improve the therapeutic effect of peripheral facial palsy at recovery stage.

[Wogonoside reverses cisplatin resistance in SGC7901/cDDP cells through inhibition of PI3K/Akt/Nrf2/ARE signaling pathway].

The aim of this study was to investigate the effect of wogonoside (WGS) on the cisplatin (cDDP) resistance in human gastric carcinoma SGC7901/cDDP cells and its possible mechanism. The drug-resistant SGC7901/cDDP cells were established by stepwise exposure to cDDP. CCK-8 assay was employed to detect the cytotoxic effect of WGS and cDDP on SGC7901/cDDP cells, and the combined effect of WGS and cDDP was analyzed by Chou-Talalay method. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). Nuclear factor erythroid-2 related factor 2 (Nrf2) gene was knocked down by using the short hairpin RNA (shRNA) approach. The protein levels of Nrf2, NAD(P)H:quinone oxidoreductase 1 (NQO1), glutathione S transferase-π (GST-π), multidrug resistance-associated protein 1 (MRP1), cleaved Capase-3, p-Akt and Akt were detected by Western blotting. The result showed that after various concentrations of WGS and/or cDDP treatment for 48 h, the cell viability was remarkably reduced in a dose-dependent manner (P < 0.05). When the inhibition rate exceeded 16%, the combination of WGS and cDDP produced a synergistic effect. The protein levels of p-Akt, Nrf2 and MRP1 in SGC7901/cDDP cells were higher than those in SGC7901 cells (P < 0.05). WGS and LY294002 (PI3K inhibitor) both remarkably decreased the phosphorylation level of Akt (P < 0.05), down-regulated the protein level of Nrf2 (P < 0.05), increased the content of ROS (P < 0.05), up-regulated the protein level of cleaved Caspase-3 (P < 0.05), and induced apoptosis (P < 0.05). Meanwhile, N-acetyl-L-cysteine (NAC) decreased apoptosis and oxidative stress reaction induced by WGS (P < 0.05). WGS and Nrf2 gene silencing both down-regulated the protein levels of NQO1, GST-π and MRP1 (P < 0.05). These results suggest that WGS may reverse cDDP resistance in SGC7901/cDDP cells through blocking the PI3K/Akt/Nrf2/ARE signaling pathway, thus enhancing the cytotoxicity of cDDP and inducing oxidative stress reaction and apoptosis.

Astragaloside IV prevents lipopolysaccharide-induced injury in H9C2 cardiomyocytes.

This study aimed to investigate the protective effects of astragaloside IV (AS IV) on lipopolysaccharide (LPS)-induced injury in H9C2 cardiomyocytes. H9C2 Cardiomyocytes were cultured with LPS (10 µg·mL(-1)) for 4 h and treated with AS IV at 50, 100, and 150 µmol·L(-1) for various durations. Cell viability was determined by MTT. The content of released TNF-α and IL-6 from cardiomyocytes were evaluated by enzyme-linked immunosorbent assay (ELISA). The levels of superoxidase dismutase (SOD), malondialdehyde (MDA), lactate dehydrogenase (LDH), and creatine phosphate kinase (CK) were measured by using commercial available kits. The mRNA and protein expression levels of NF-κB p65 were measured by RT-PCR and Western blotting, respectively. And the NF-κB p65 activity was measured by ELISA. Our results demonstrated that AS IV at 50, 100, and 150 µmol·L(-1) markedly inhibited the release of TNF-α and IL-6 and decreased NF-κB expression, compared with the model group. Moreover, the improved SOD activity and decreased MDA, LDH and CK levels were detected after AS IV treatment. In summary, AS IV could increase the activities of antioxidant enzymes, inhibite lipid peroxidation, and down-regulate the inflammatory mediators involved in the inflammatory responses. These results demonstrated that AS IV could prevent LPS-induced injury in cardiomyocytes.

[Autophagy and prostate cancer].

The role of autophagy is known to be highly complex and context-dependent, and may be characterized as both tumor suppression and tumor promotion in some tumors, such as breast cancer and prostate cancer. This review outlines recent advances in the studies of the involvement of autophagy in the development, progression and treatment of prostate cancer, focusing on autophagy modulation during androgen deprivation, with a special discussion on the regulatory effect of androgens on the autophagy of prostate cancer cells. A critical evaluation and analysis of the studies suggests that autophagy inhibition combined with androgen deprivation therapy is a promising approach to the treatment of prostate cancer.

Reversal of multidrug resistance by 5,5'-dimethoxylariciresinol-4-O-ß-D-glucoside in doxorubicin-resistant human leukemia K562/DOX.

OBJECTIVE: The objective of this study was to investigate the reversal effects of 5,5'-dimethoxylariciresinol-4'-O-ß-D-glucoside (DMAG) extracted from traditional Chinese medicines Mahonia on multidrug resistance (MDR) of human leukemia cells to chemotherapeutic agents. MATERIALS AND METHODS: MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was performed to determine the effect of DMAG on doxorubicin sensitivity to K562/DOX cells. Propidium iodide /Hoechst 33342 double staining assay was used to investigate the effect of DMAG on doxorubicin-induced cellular apoptosis. Intracellular accumulation of doxorubicin and rhodamine 123 assay were performed to evaluate the effect of DMAG on drugs efflux activity of P-glycoprotein. RESULTS: DMAG significantly enhanced the doxorubicin cytotoxicity to K562/DOX cells. In the presence of 1.0 µM of DMAG, the IC50 of doxorubicin decreased from 34.93 ± 1.37 µM to 12.51 ± 1.28 µM. DMAG of 1.0 µM significantly enhanced doxorubicin-induced cell apoptosis in K562/DOX cells and the enhancement was time-dependent. A significant increase in accumulation of doxorubicin in the presence of DMAG was observed. After treatment of the K562/DOX cells for 1 h with 15.0 µM doxorubicin alone, the fluorescence intensity was 33093.12. With the addition of 1.0 µM of DMAG, the fluorescence intensity of doxorubicin was 2.3-fold higher. A significant increase of accumulation of rhodamine 123 in the presence of DMAG was also observed. With the addition of 1.0 µM of DMAG, the fluorescence intensity was increased by 49.11% compared with rhodamine 123 alone. CONCLUSION: DMAG was shown to effectively enhance chemosensitivity of resistant cells, which makes it might be a suitable candidate for potential MDR-reversing agents.

Astragaloside IV stimulates angiogenesis and increases nitric oxide accumulation via JAK2/STAT3 and ERK1/2 pathway.

Astragaloside IV (AS-IV), one of the major active constituents of Astragalus membranaceus in Traditional Chinese Medicine, has been widely used to treat ischemic diseases. However, the potential mechanism is this action is unclear. In this study, we tested the hypothesis that AS-IV might promote angiogenesis through multiple signaling pathways. Our data indicate that AS-IV treatment promotes umbilical vein endothelial cells (HUVEC) proliferation, migration, and tube formation. AS-IV treatment also activates JAK2/STAT3 and ERK1/2 signaling pathways, and up-regulates endothelial nitric oxide synthase (eNOS) expression and nitric oxide (NO) production. AS-IV-induced angiogenesis in HUVECs is significantly blocked by specific kinase inhibitors. Our study indicated that AS-IV is a key regulator of NO and angiogenesis through the JAK2/STAT3 and ERK1/2 pathways, which provides a mechanistic basis for the potential use of this compound in the treatment of clinical ischemic diseases.

[Clinical observation on acupuncture at Zhaohai (KI 6) and Shenmai (BL 62) for treatment of insomnia].

OBJECTIVE: To observe therapeutic effect of acupuncture at Zhaohai (KI 6) and Shenmai (BL 62) on insomnia. METHODS: Seventy-eight cases of insomnia were randomly divided into a treatment group of 40 cases and a control group of 38 cases. The treatment group were treated with acupuncture at Zhaohai (KI 6) using reinforcing method and at Shenmai (BL 62) using reducing method, combined with acupuncture at acupoints selected according to syndrome differentiation. The control group were treated with acupuncture at acupoints selected according to syndrome differentiation. Their therapeutic effects were compared. RESULTS: The cured rate and the total effective rate were 62.5% and 97.5% in the treatment group, and 31.6% and 68.4% in the control group, respectively, with significant difference between the two groups (P < 0.01). CONCLUSION: Acupuncture at Zhaohai (KI 6) and Shenmai (BL 62) has a better therapeutic effect on insomnia.