Efficacy of Yushen Tongluo Granule Combined with Clomiphene Citrate for Anovulatory Infertility: A Double-Blind, Randomized, Placebo-Controlled Clinical Trial.

OBJECTIVE: To evaluate the efficacy and safety of Yushen Tongluo Granule (YSTLG) combined with clomiphene citrate (CC) in the treatment of anovulatory infertility. METHODS: This randomized, double-blinded, placebo-controlled clinical trial was carried out in the Department of Obstetrics and Gynecology and the Department of Traditional Chinese Medicine (TCM). During the 3 menstrual cycle intervention periods, all subjects received 50 mg/day CC from day 5 until day 9 of the menstruation. If no ovulation, the amount of CC per cycle increased 50 mg/day until 150 mg/day. Participants in the experimental group received YSTLG, while participants in the control group received YSTLG placebo. The granules were orally taken from the end of menstruation until ovulation. When one leading follicle attained a diameter of 18 mm or more, 5000 U human chorionic gonadotropin (hCG) was given intramuscularly. The primary outcome measure was the ovulation rate, and follicular development was monitored by transvaginal ultrasound on the 10th day of the cycles until ovulation. Secondary outcome measures including the overall curative effect, endometrial thickness, and pregnancy outcomes were also compared between the two groups. RESULTS: The ovulation rate in the experimental group was higher than that in the control group (P < 0.05). Compared with the control group, the overall curative effect of the experimental group was better than that of the control group (P < 0.05), and the endometrial thickness in the ovulation phase was significantly thicker than that in the control group (P < 0.01). There was no significant difference in pregnancy rate and miscarriage rate between the experimental group and control group (P > 0.05). CONCLUSION: The combined YSTLG and CC used to treat anovulatory infertility can improve the ovulation rate without affecting endometrial thickness, which is efficacious and safe.

Propofol induces ROS-mediated intrinsic apoptosis and migration in triple-negative breast cancer cells.

Propofol is widely applied in general anesthesia owing to its short effect and rapid recovery. Apart from its anesthetic advantages, propofol has also been observed to inhibit the growth of several types of cancer cells. Breast cancer is the most diagnosed cancer in females worldwide and triple negative breast cancer (TNBC) constitutes 15-20% of all breast cancer cases. TNBC is characterized by a high recurrence rate, which is associated with its high mortality rate. The present study aimed to evaluate apoptosis in MDA-MB-468 cells treated with propofol. The Cell Counting Kit-8 assay was used to assess proliferation in cells treated with different concentrations of propofol. In addition, Annexin V-FITC was used to detect apoptosis. Furthermore, the generation of reactive oxygen species (ROS) was examined. The relative expression of proteins in the intrinsic apoptosis pathway, such as Bak, Bax, Bcl-2, Cytochrome c, apoptotic peptidase-activating factor 1 (Apaf-1), Caspase 3 and Caspase 9, were calculated relative to GAPDH with western blot analysis. A wound healing assay was performed to examine the effect of propofol on MDA-MB-468 cell migration. The present study revealed that propofol inhibited the proliferation and increased the level of ROS in MDA-MB-468 cells. The expression levels of Cytochrome c, Apaf-1, Bax, Bak and cleaved Caspase 3/9 were upregulated compared with GAPDH. The level of Bcl-2 protein was upregulated by propofol at a concentration of 5 µM and downregulated at concentrations of 10 and 20 µM. In the wound-healing assay, propofol reduced the scratch distance and area. Taken together, the results of the present study suggested that propofol may induce ROS-mediated intrinsic apoptosis and promote migration in TNBC cells.

The active fraction of Garcinia yunnanensis suppresses the progression of colorectal carcinoma by interfering with tumorassociated macrophage-associated M2 macrophage polarization in vivo and in vitro.

Colorectal cancer (CRC) is the third most common solid tumor worldwide and has shown resistance to several immunotherapies, particularly immune checkpoint blockade therapy, which is effective in many other types of cancer. Our previous studies indicated that the active fraction of Garcinia yunnanensis (YTE-17), had potent anticancer activities by regulating multiple signaling pathways. However, knowledge regarding the mechanism and effect of YTE-17 in the prevention of CRC is limited. This study tested the effects of YTE-17 on colon cancer development in vivo by using two murine models: the carcigenic azoxymethane/dextran sulfate sodium (AOM/DSS)-induced CRC model and a genetically induced model using ApcMin/+ mice. Here, the tumor load, tumor number, histology, and even some oncogenes were used to evaluate the effect of YTE-17. The intragastric administration of YTE-17 for 12 weeks significantly decreased CRC incidence, tumor number and size, immunity, and some tumor-associated macrophage (TAM) markers, including CD206, Arg-1, IL-10, and TGF-ß. Importantly, the macrophages depletion by clodronate (CEL) also played a role in reducing the tumor burden and inhibiting tumor development, which were not affected by YTE-17 in the ApcMin/+ mice. Moreover, the YTE-17 treatment attenuated CRC cell growth in a co-culture system in the presence of macrophages. Consistently, YTE-17 effectively reduced the tumor burden and macrophage infiltration and enhanced immunity in the AOM/DSS and ApcMin/+ colon tumor models. Altogether, we demonstrate that macrophages in the microenvironment may contribute to the development and progression of CRC cells and propose YTE-17 as a new potential drug option for the treatment of CRC.

MALAT1: A long non-coding RNA highly associated with human cancers.

Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), a well-known lncRNA associated with numerous diseases, particularly cancer, has received increased attention. The expression of MALAT1 was determined to be upregulated in numerous types of tumors and MALAT1 exhibited effects on tumor cell proliferation, migration, invasion and apoptosis. The abnormal expression of MALAT1 was identified in almost in every organ of the digestive system. MALAT1 performed an important role in the pathological alterations of organs that are associated with sex hormones and several reproductive system cancers. MALAT1 participates in molecular pathways. In the clinical application of MALAT1, MALAT1 was considered as a potential biomarker for the diagnosis and prediction of cancers, and may also serve as therapeutic target for treatment of specific tumors. This review summarizes the abnormal expression of MALAT1 in cancer, its significant effect on the primary features of cancer, as well as the underlying molecular mechanisms of MALAT1 in various cancers. According to studies on MALAT1, we introduce the upstream and downstream substances associated with the function of MALAT1. These reviewed studies promote the clinical application of MALAT1 in the aspect of diagnosis and treatment of different cancers, and may help point out new study directions for MALAT1.

Zhi Zhen Fang formula reverses Hedgehog pathway mediated multidrug resistance in colorectal cancer.

Zhi-Zhen-Fang (ZZR), a Traditional Chinese Medicine (TCM) formula, has been clinically used in China to treat drug-resistant colorectal cancer (CRC) patients as an adjuvant. In this study, the efficacy of ZZR in suppressing multidrug resistance (MDR) on CRC was evaluated in vitro and in vivo. We observed that ZZR enhanced the sensitivity of chemotherapeutic drugs and induced apoptosis in a dose- and time-dependent mannner in CRC MDR cells. Interestingly, signaling of Hedgehog pathway, particularly Gli1, was also inhibited by ZZR. This effect of ZZR in reversing drug resistance and suppressing Gli1 was attenuated by a Hedgehog activator (SAG). Furthermore, ZZR inhibited MDR CRC tumor growth in a xenograft mouse model as well as downregulated Gli1 levels. This study provided the first direct evidence demonstrating ZZR can attenuate MDR by repressing Hedgehog signaling in human CRC.

miR-30a acts as a tumor suppressor by double-targeting COX-2 and BCL9 in H. pylori gastric cancer models.

Helicobacter pylori (H. pylori) is one of the most important factors that affect the development of gastric cancer, and its mechanism remains un-elucidated. Our present study found that, miR-30a is crucial for regulating the growth and migration of H. pylori infected gastric cancer in vitro by targeting COX-2 and BCL9. In details, double-stranded miR-30a precursor produced two single-stranded and matured miRNAs including miR-30a-3p and miR-30a-5p, which played significant biological functions in two different manners. First, miR-30a-3p inhibited COX-2 expression and regulated nuclear translocation of ß-catenin, and second, miR-30a-5p targeted BCL9 to regulate TCF/LEF promoter activity followed by affecting ß-catenin downstream target gene expression. In vivo, miR-30a knockout mice were successfully achieved using CRISPR/Cas9 gene editing technology. Compared with H. pylori-infected wild-type mice, H. pylori-infected miR-30a knockout mice showed increased incidence of chronic gastritis, chronic atrophic gastritis, atypical hyperplasia, and other precancerous lesions or adenocarcinoma manifestations in the antral or gastric mucosa of mice, as well as regulation of genes closely associated with tumor development. Taken together, miR-30a acts as a tumor suppressor by double-targeting COX-2 and BCL9, and significantly affects the development of H. pylori-induced gastric cancer, shedding new light on the mechanisms underlying H. pylori-associated gastric cancer.

Salvianolic acid B reverses multidrug resistance in HCT­8/VCR human colorectal cancer cells by increasing ROS levels.

Salvianolic acid B (SalB) a water­soluble phenolic compound, extracted from Salvia miltiorrhiza, has previously been demonstrated to reverse tumor multidrug resistance (MDR), including in colorectal cancer. Reactive oxygen species (ROS) are oxygen radicals generated during aerobic metabolism (superoxide and hydroxyl radicals) and superoxide easily generating free radicals (H2O2). The concept that increased ROS levels can lead to augmented tumor cell­sensitivity to chemotherapy drugs has become notable. The aim of the present study was to elucidate the role of ROS in mediating the effect of SalB on drug resistance and the correlation with drug resistance­associated protein, P­glycoprotein (P­gp), and apoptosis­associated proteins, B­cell lymphoma 2 (Bcl­2) and Bcl­2­associated X (Bax). In the current study, through utilizing the multidrug resistant colorectal cancer cell line, HCT­8/VCR, it was demonstrate that SalB reversed MDR in HCT­8/VCR. In addition, SalB significantly increased ROS levels, which may have accelerated the apoptosis of HCT­8/VCR cells by downregulating Bcl­2 and increasing Bax protein expression. Furthermore the increased intracellular ROS levels may have inhibited P­gp expression at the gene and protein levels. In conclusion, the data of the current study demonstrate that SalB reversed MDR in HCT­8/VCR cells, and the effect is associated with increased ROS levels, which may downregulate P­gp expression and promote tumor cell apoptosis, which in turn increases the sensitivity of drug­resistant cells to chemotherapy drugs.

[Medicated serum prepared with Chinese herbal medicine Zhizhen Recipe down-regulates activity of nuclear factor-κB and expression of P-glycoprotein in human colorectal cancer multidrug-resistant cell line HCT-8/VCR].

OBJECTIVE: To investigate the effects of medicated serum prepared with Chinese herbal medicine Zhizhen Recipe (ZZR) on activity of nuclear factor-κB (NF-κB) and expression and function of P-glycoprotein (P-gp) in human colorectal cancer multidrug-resistant cell line HCT-8/VCR. METHODS: The multidrug resistance of HCT-8/VCR cells was detected by cell counting kit-8 method, and the experimental concentrations of ZZR-medicated serum were determined by the same way. HCT-8 and HCT-8/VCR cells were treated with ZZR-medicated serum of medium dose for 24 h. The activity of NF-κB was determined by enzyme-linked immunosorbent assay. The intracellular distribution of P-gp was detected by laser scanning confocal microscopy, and the mean fluorescence intensity of rhodamine 123 was detected by flow cytometry. RESULTS: ZZR-medicated sera with volume fraction of 8%, 16% and 32% of medium dose were confirmed as the experimental sera. Compared with the untreated group, NF-κB activities of the ZZR-medicated serum groups (ZZR-medicated serum with volume fraction of 8%, 16% and 32% of medium dose) were obviously down-regulated (P<0.01), which had a negative correlation with the concentrations. After interfering HCT-8/VCR with ZZR-medicated serum of different concentrations for 24 h, P-gp in HCT-8/VCR transmitted gradually from cell membrane to cytoplasm and nuclei. Nuclei became pyknotic and cracking. Compared with the untreated group, the mean fluorescence intensities of ZZR-medicated serum groups declined with concentration gradients (P<0.01). The efflux of intracellular rhodamine 123 decreased, the wave crest shifted to right, and the intracellular fluorescence intensity strengthened (P<0.01). CONCLUSION: ZZR-medicated sera of experimental concentrations down-regulate activity of NF-κB and expression and function of P-gp in human colorectal cancer multidrug-resistant cell line HCT-8/VCR and the effect is related to the concentrations.

[Clinical study of wind-warm and pulmonary heat syndrome treated with integrated traditional Chinese and Western medicine].

OBJECTIVE: To investigate the clinical effectiveness of Toubiao Qingfei (expelling exterior evil and clearing lung) Decoction (TBQFD) on wind-warm and pulmonary heat syndrome. METHODS: Forty-six subjects were randomized into treatment group and control group. Patients in the control group were treated with Western medicines, while patients in the treatment group were treated with Western medicines and TBQFD. The therapeutic effects and improvement of symptoms in both groups were observed. RESULTS: The durations of fever, cough and absorption of pulmonary inflammatory focus in the treatment group were 1.52, 3.52 and 6.25 days respectively, which were significantly shorter than those in the control group (P<0.05). CONCLUSION: TBQFD can improve the symptoms, such as fever and cough, and can promote the absorption of infection and shorten the clinical course of wind-warm and pulmonary heat syndrome.

[A study on relationship of purple tongue and its regional blood flow].

OBJECTIVE: To study the relationship of purple tongue and its regional blood flow. METHODS: Laser-Doppler flow meter was employed to test the volume and velocity of blood flow at different sites of purple tongue (56 cases) and normal tongue (54 cases), and the test results were analyzed. RESULTS: The volume of blood flow of purple tongue was significantly lower than that of the normal tongue (P<0.01), while the velocity was significantly higher (P<0.01). The volume and velocity of blood flow at different sites of the tongue were similar without significant difference. CONCLUSION: The volume of blood flow of purple tongue is decreased and the velocity increased. Purple tongue may be caused by dysfunction of the microcirculation of the tongue.