Phylogeny, character evolution, and biogeography of the fern genus Bolbitis (Dryopteridaceae).

Bolbitis is a pantropical fern genus of Dryopteridaceae with ca. 80 species mainly in tropical Asia. Earlier studies confirmed the monophyly of Bolbitis when Mickelia is excluded and identified three major clades in Bolbitis. However, earlier studies are based on relatively small sampling and the majority of Asian species are not sampled. In this study, DNA sequences of three plastid markers of 169 accessions representing ca. 68 (85 % of total) species of Bolbitis in nine out of the 10 series recognized by Hennipman (1977), and 54 accessions representing the five remaining bolbitidoid genera are used to infer a global phylogeny with a focus on Asian species. The major results include: (1) Bolbitis is strongly supported as monophyletic; (2) species of Bolbitis are resolved into four major clades and their relationships are: the Malagasy/Mascarene clade is sister to the rest, followed by the African clade which is sister to the American clade + the Asian clade; (3) six well-supported subclades are identified in the most speciose Asian clade; (4) the free-veined Egenolfia is embedded in Bolbitis and is paraphyletic in relation to species with anastomosing venation; (5) three series sensu Hennipman (1977), B. ser. Alienae, B. ser. Egenolfianae, and B. ser. Heteroclitae, are paraphyletic or polyphyletic; (6) evolution of six morphological characters is analyzed and free venation is found to have evolved from anastomosing venation and reversed to free venation in Bolbitis; and (7) biogeographical implications are drawn and it is shown that a single recent dispersal from Asia resulted in continental disjunction of closely related ferns of Bolbitis between Africa and America.

Genome size evolution of the extant lycophytes and ferns.

Ferns and lycophytes have remarkably large genomes. However, little is known about how their genome size evolved in fern lineages. To explore the origins and evolution of chromosome numbers and genome size in ferns, we used flow cytometry to measure the genomes of 240 species (255 samples) of extant ferns and lycophytes comprising 27 families and 72 genera, of which 228 species (242 samples) represent new reports. We analyzed correlations among genome size, spore size, chromosomal features, phylogeny, and habitat type preference within a phylogenetic framework. We also applied ANOVA and multinomial logistic regression analysis to preference of habitat type and genome size. Using the phylogeny, we conducted ancestral character reconstruction for habitat types and tested whether genome size changes simultaneously with shifts in habitat preference. We found that 2C values had weak phylogenetic signal, whereas the base number of chromosomes (x) had a strong phylogenetic signal. Furthermore, our analyses revealed a positive correlation between genome size and chromosome traits, indicating that the base number of chromosomes (x), chromosome size, and polyploidization may be primary contributors to genome expansion in ferns and lycophytes. Genome sizes in different habitat types varied significantly and were significantly correlated with habitat types; specifically, multinomial logistic regression indicated that species with larger 2C values were more likely to be epiphytes. Terrestrial habitat is inferred to be ancestral for both extant ferns and lycophytes, whereas transitions to other habitat types occurred as the major clades emerged. Shifts in habitat types appear be followed by periods of genomic stability. Based on these results, we inferred that habitat type changes and multiple whole-genome duplications have contributed to the formation of large genomes of ferns and their allies during their evolutionary history.

Qigong for women with breast cancer: An updated systematic review and meta-analysis.

OBJECTIVE: The purpose of this review was to evaluate the effectiveness of Qigong in improving the quality of life and relieving fatigue, sleep disturbance, and cancer-related emotional disturbances (distress, depression, and anxiety) in women with breast cancer. METHODS: The PubMed, Cochrane Central Register of Controlled Trials, Web of Science, Sinomed, Wanfang, VIP, and China National Knowledge Infrastructure databases were searched from their inceptions to March 2020 for controlled clinical trials. Two reviewers selected relevant trials that assessed the benefit of Qigong for breast cancer patients independently. A methodological quality assessment was conducted according to the criteria of the 12 Cochrane Back Review Group for risk of bias independently. A meta-analysis was performed by Review Manager 5.3. RESULTS: This review consisted of 17 trials, in which 1236 cases were enrolled. The quality of the included trials was generally low, as only five of them were rated high quality. The results showed significant effectiveness of Qigong on quality of life (n = 950, standardized mean difference (SMD), 0.65, 95 % confidence interval (CI) 0.23-1.08, P =  0.002). Depression (n = 540, SMD = -0.32, 95 % CI -0.59 to -0.04, P =  0.02) and anxiety (n = 439, SMD = -0.71, 95 % CI -1.32 to -0.10, P =  0.02) were also significantly relieved in the Qigong group. There was no significant benefit on fatigue (n = 401, SMD = -0.32, 95 % CI  0.71 to 0.07, P = 0.11) or sleep disturbance relief compared to that observed in the control group (n = 298, SMD = -0.11, 95 % CI  0.74 to 0.52, P = 0.73). CONCLUSION: This review shows that Qigong is beneficial for improving quality of lifeand relieving depression and anxiety; thus, Qigong should be encouraged in women with breast cancer.

[Correlation Study between Syndrome Types of Chinese Medicine and Molecular Subtyping of Breast Cancer in Consolidate Period].

Objective To observe the correlation between syndrome types of Chinese medicine (CM) and molecular subtyping of breast cancer in consolidation period. Methods Multivariate statistical analysis was applied in this research. Totally 612 breast cancer patients in consolidation period were as- signed to 4 syndrome types of CM [qi deficiency syndrome (22. 22% , 136/612) , Gan-qi stagnation syn- drome (21.73%, 133/612), Gan-Shen yin deficiency syndrome (20.10%, 123/612), Shen deficiency blood stasis syndrome (35. 95%, 220/612) ]. The correlation between each syndrome type and each molecular subtyping was respectively analyzed using Χ² (R x C table) test. Results Through statistical a- nalysis and expert consultation, results showed that cluster four types in consolidate period of breast cancer were compatible. Each syndrome type was correlated with each molecular subtyping in 612 breast cancer patients in consolidation period. Luminal type A was correlated with Gan-qi stagnation syndrome (P <0. 05). Luminal type B and triple negative type were correlated with qi deficiency syndrome and Gan- Shen yin deficiency syndrome (P <0. 05). But each syndrome type was not obviously correlated with dis- ease course. There was no obvious correlation between molecular subtyping and age/disease course (P >0. 05). Conclusion Luminal type A occupied the highest ratio in Gan-qi stagnation syndrome, with relatively better prognosis.

Yiqi formula enhances the antitumor effects of erlotinib for treatment of triple-negative breast cancer xenografts.

Yiqi formula (YF), a traditional herbal prescription, has long been used to treat triple-negative breast cancer (TNBC) patients. The present study aims to investigate the effects and the related mechanism of YF for treatment of TNBC xenografts. MDA-MB-231 (human TNBC) cells were subcutaneously injected into the second mammary fat pad of 40 female nude mice, which were divided into four groups: control, erlotinib (an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor), YF, and combination (YF plus erlotinib). All treatments were administered orally for 30 days. Inhibition rate of tumor weight by erlotinib, YF, and the combination was 26.47%, 17.24%, and 39.15%, respectively. Western blotting showed that YF, erlotinib, and the combination downregulated p-EGFR (P < 0.01) and p-Akt1 (pT308) (P < 0.05) and upregulated PTEN compared with control, and the combination was more efficacious than erlotinib alone (P < 0.05). Similar results were detected by immunohistochemistry. Real-time quantitative PCR showed that YF, erlotinib, and the combination increased PTEN mRNA (P < 0.05, P < 0.01) compared with control, and the combination was more efficacious than erlotinib alone (P < 0.05). In conclusion, YF can regulate the main components of the PI3K/Akt pathway in TNBC xenografts. When YF was used in combination with erlotinib, it enhanced the antitumor effects of erlotinib on TNBC xenografts. These findings suggest that YF is suitable to use for the treatment of TNBC patients.

[Effects of Astragalus polysaccharide on proliferation and Akt phosphorylation of the basal-like breast cancer cell line].

OBJECTIVE: To investigate the effects of Astragalus polysaccharide (APS) on proliferation of basal-like breast cancer cell line MDA-MB-468 cells and Akt phosphorylation in MDA-MB-468 cells. METHODS: APS at different concentrations was used to culture MDA-MB-468 cells for different time periods, and then proliferation of MDA-MB-468 cells was assayed using methyl thiazolyl tetrazolium (MTT) assay to determine the time- and dose-dependent effects of APS. For observing the effects of APS on phosphor-Akt (p-Akt), in-cell Western blot method was used after 1, 2, 4 and 7 d of culture in APS to detect protein expressions of p-Akt (Thr308) and p-Akt (Ser473). Protein levels of the key targets in p53/murine double minute 2 (MDM2) signaling pathway, such as p53, MDM2 and phosphatase and tensin homolog deleted on chromosome ten (PTEN) were also detected. After PTEN gene was silenced by small interfering RNA (siRNA) in MDA-MB-468 cells, expressions of p-Akt (Thr308 and Ser473) were assayed by the in-cell Western blot method after 2 d of APS treatment. RESULTS: APS at 1 and 0.5 mg/mL concentrations effectively inhibited the proliferation of MDA-MB-468 cells and was used in subsequent tests. Compared with the control group, APS decreased the protein expression of p-Akt (Thr308) in MDA-MB-468 cells after 1-, 2-, 4- and 7-day culture, and also decreased the protein expression of p-Akt (Ser473) and up-regulated the protein expression of MDM2 in MDA-MB-468 cells after 1- and 2-day culture. Expressions of p53 and PTEN were up-regulated after 7 d of APS culture. After silencing PTEN gene by siRNA, APS could not mediate Akt phosphorylation. CONCLUSION: APS can inhibit proliferation of basal-like breast cancer cell line MDA-MB-468, and down-regulate the expression of Akt phosphorylation. The antiproliferation mechanisms may be related to its effects of up-regulating the expressions of p53 and PTEN by regulating p53/MDM2 positive and negative feedback loops.

[Study protocol for a self-controlled case study to evaluate the safety and standardization for external application of Chinese medicine Jiuyi Powder].

BACKGROUND: As the main medicinal powder for drawing out pus and removing necrotic tissue in external therapies of traditional Chinese surgery, Sheng Powder has made great contributions to the treatment of inflammatory wounds and has the unique bactericidal and decay-discharging function that can not be replaced by antibiotics. However, Sheng Powder has toxicity because it contains mercury. So far, there is no clinical research on the standards of dose and usage of Sheng Powder and there is a lack of objective and quantitative criteria for operating standards and monitoring of toxicity and side effects. Therefore, the authors choose Jiuyi Powder, one of the most commonly used Sheng Powder, to evaluate the safety of its external use, and form a standardization program for clinical implementation. METHODS AND DESIGN: This study is a nonrandomized, nonblinded, self-controlled case study. There will be two key stages in the study. In stage one, 10 patients with plasma cell mastitis will be enrolled. The patients will receive continuous external application of Jiuyi Powder with a fixed dose (160 mg/d). Blood mercury and urine mercury levels will be dynamically tested at different time intervals, and the observation of pharmacokinetic parameters will be conducted after Jiuyi Powder has been absorbed by the surface of the wound. In stage two, 30 patients with each of the four conditions including postoperative wounds of head or facial surgeries, plasma cell mastitis, anal fistula and chronic absorption ulcer of the shank will be enrolled, respectively. According to the dose of 1.5 mg/cm(2), Jiuyi Powder will be externally applied to the wound surface for 14 d. On the basis of the first-phase study, the test time will be chosen and the toxicity outcome will be detected to evaluate the safety of external application of Jiuyi Powder and to establish recommendations for standardized clinical use. DISCUSSION: The purpose of this study is to evaluate the safety of the external application of Jiuyi Powder through a two-stage study. The pharmacokinetic parameters of external application of Jiuyi Powder at the clinical dose range, the changes in blood and urinary mercury levels and related safety indexes will be observed on different wounds located in different positions externally. The evaluation of whether Jiuyi Powder could be a routine medication for drawing out pus and removing necrotic tissue in inflammatory and refractory wounds will be stated. TRIAL REGISTRATION NUMBER: ChiCTR-TNC-11001366.

[Effects of Astragalus injection and its ingredients on proliferation and Akt phosphorylation of breast cancer cell lines].

OBJECTIVE: To observe the effects of Astragalus injection, astragaloside IV and formononetin on proliferation and Akt phosphorylation of basal-like human breast carcinoma cell lines MDA-MB-468 and MDA-MB-231. METHODS: The effects of different concentrations of Astragalus injection, astragaloside IV and formononetin on proliferation of breast cancer cell lines were assayed by methyl thiazolyl tetrazolium (MTT) assay, and their effects on phospho-Akt were assayed by in-cell Western blot method. RESULTS: The results of the MTT assay showed that the best concentrations of Astragalus injection, astragaloside IV, formononetin and astragaloside IV plus formononetin were 1 g/mL, 80 microg/mL, 40 microg/mL and 10 microg/mL plus 40 microg/mL respectively. After 1- or 2-day culture, Astragalus injection, astragaloside IV, formononetin and astragaloside IV plus formononetin decreased the expressions of p-Akt (Thr 308) and p-Akt (Ser 473) in MDA-MB-468 cells. Formononetin and astragaloside IV plus formononetin down-regulated the expression of p-Akt (Thr 308) protein in MDA-MB-231 cells after 1- and 2-day culture, but had no effects on the expression of p-Akt (Ser 473) protein in MDA-MB-231 cells. CONCLUSION: Astragalus injection, astragaloside IV and formononetin can inhibit proliferation of breast cancer cell lines MDA-MB-468 and MDA-MB-231, and the antiproliferation effects vary according to their concentrations. And the antiproliferation mechanisms may be related to their down-regulation effects on Akt phosphorylation.

[Study on surface infrared radiation spectrums of danzhong (CV 17) in the patient of hyperplasia of mammary glands].

OBJECTIVE: To compare differences of surface infrared radiation spectrums between Danzhong (CV 17) A high-sensitive PHE and non-acupoint control point in the patient of hyperplasia of mammary glands. METHODS: 201 surface infrared spectrograph was used to detect infrared radiation spectrums of Danzhong (CV 17) and non-acupoint control point at 1.5-16.0 microm wave band in the patient of hyperplasia of mammary glands. RESULTS: The shape of the infrared spectrums of Danzhong (CV 17) was similar to that of the non-acupoint control point, but with lower radiation intensity. Of the 59 wavelength spots detected, 13 (6.75-8.25 microm, 9.00 microm, 9.25 microm, 9.75 microm, 13.25-13.75 microm) had significant differences in infrared radiation intensity between Danzhong (CV 17) and non-acupoint control point (P<0.05). CONCLUSION: The intensity of infrared radition of Danzhong (CV 17) is lower and Qimen (LR 14) is higher than that of the non-acupoint control point in the patient with hyperplasia of mammary glands, showing that different channels are at different states of deficiency and excess under pathological condition of hyperplasia of mammary glands.

[Research progress on basal-like breast cancer].

Breast cancer is the most common cancer in women all around the world. Presently, the prognosis of breast cancer is predicted based on histopathologic tumor characteristics. Molecular markers could help to predict the prognosis and treatment response in breast cancer. According to gene expression profile, invasive breast cancers are classified into four subtypes (luminal A, luminal B, HER2 overexpressing and basal-like) and associated with different prognosis. The basal-like subtype is associated with poor prognosis. This article simply introduced the distribution of molecular subtypes, summed up recent researches on the characteristics and prognosis of basal-like phenotype.

[Effects of Astragalus injection on proliferation of basal-like breast cancer cell line MDA-MB-468].

OBJECTIVE: To investigate the effects of Astragalus injection (AI) on basal-like breast cancer cell line MDA-MB-468 and murine bone marrow stromal stem cells (mMSCs). METHODS: MDA-MB-468 cells and primary cultured mMSCs were treated by different concentrations of AI, and with untreated MDA-MB-468 cells as blank control. The morphology of cells was observed by phase-contrast inverted microscope and transmission electron microscopy. Cytotoxic effects of AI on MDA-MB-468 cells and mMSCs were evaluated by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis of MDA-MB-468 cells induced by AI were measured by flow cytometry. Lactate dehydrogenase (LDH) activity in supernatants was measured by enzymatic colorimetric method. The expressions of epidermal growth factor receptor (EGFR) and p53 protein in MDA-MB-468 cells were evaluated by streptavidin-biotin-peroxidase complex method. RESULTS: A time-dependent cytotoxic effect of 1 g/ml AI was observed in MDA-MB-468 cells. 1 g/ml AI also had cytotoxic effect on mMSCs, but its effect was not better than cisplatin. 0.1 g/ml AI could promote the proliferation of mMSCs. Different concentrations of AI could all induce the apoptosis of MDA-MB-468 cells. There was no significant difference in LDH activity in the supernatants between blank control group and AI-treated and cisplatin-treated groups. AI could down-regulate the expressions of EGFR and p53 protein. CONCLUSION: The effects of AI on MDA-MB-468 cells and mMSCs are related to the concentration of AI, and its mechanism of inhibiting the proliferation of MDA-MB-468 cells may be due to down-regulation of the expressions of EGFR and p53 protein.

Effect of runing II on the growth and metastasis of transplanted tumor in mammary cancer-bearing mice and its mechanism.

OBJECTIVE: To study the effect of Runing II (a Chinese herbal preparation for mammary cancer) on the growth and metastasis of transplanted tumor of mammary cancer MA-891-bearing TA2 mice and its mechanism. METHODS: The model of mammary cancer MA-891 cell strain transplanted tumor of TA2 mice with lung metastasis were developed to observe the effect of Runing II on the growth and metastasis of the transplanted tumor. The immunohistochemical method and image analysis were adopted to detect the levels of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR), and micro-vessel count (MVC) and micro-vessel area (MVA). RESULTS: In the Runing II group, the tumor weight inhibition rate and the lung metastasis inhibition rate were 37.3% and 65.4% respectively, the tumor growth and lung metastasis were obviously inhibited; And the levels of VEGF and VEGFR, MVC and MVA were significantly decreased as compared with those in the tumor-bearing control group (P<0.05). CONCLUSION: The Chinese herbal preparation Running II can inhibit the metastasis of tumor through inhibiting the angiogenesis, and the mechanism is possibly related with down-regulation of VEGF and VEGFR expression.

Bis(2,2'-bipyridine-κN,N')(croconato-κO,O')nickel(II).

The title compound, [Ni(C(5)O(5))(C(10)H(8)N(2))(2)], lies across a crystallographic twofold axis, around which two 2,2'-bipyridine (2,2'-bipy) ligands are arranged in a propeller manner. The local geometry of the NiN(4)O(2) coordination core basically adopts an octa-hedral geometry. The mol-ecular twofold axis is along the direction of the mol-ecular dipole moment, and the complex is packed with its dipole moment alternately along the +b and -b directions. The crystal structure is stabilized by inter-molecular C-H⋯O hydrogen bonds.

(Croconato-kappa2O,O')bis(1,10-phenanthroline-kappa2N,N')cobalt(II), and the nickel(II) and copper(II) analogues.

The title complexes, [M(C5O5)(C12H8N2)2], with M = Co(II), Ni(II) and Cu(II), all lie across twofold rotation axes, around which two 1,10-phenanthroline ligands are arranged in a chiral propeller manner. The Co(II) and Ni(II) complexes are isostructural, with octahedral coordination geometry, while the local geometry of the Cu(II) complex is severely distorted from octahedral.

[Effects of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture].

OBJECTIVE: To evaluate the effects of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture. METHODS: The normal human mammary epithelial cells were dissociated by digestion with collagenase type I. The morphological identification of normal human mammary epithelial cells in primary culture was determined under inverted phase contrast microscope and transmission electron microscope. The estradiol and progesterone were added to culture medium to induce the proliferation of normal human mammary epithelial cells in primary culture, and the different doses of Chinese herbal medicine for warming kidney and tamoxifen were also added into the culture medium. The MTT assay was used to evaluate the proliferation of normal human mammary epithelial cells in primary culture under the different conditions of interventions. RESULTS: The normal human mammary epithelial cells in primary culture presented typically morphological features of normal human mammary epithelial cells in vitro. The proliferation of normal human mammary epithelial cells in primary culture was significantly improved by intervention of estradiol and progesterone both with concentration of 1x10(-5) g/L. The high-dose Chinese herbal medicine for warming kidney could obviously promote the proliferation of normal human mammary epithelial cells in primary culture, while the proliferation was obviously inhibited by low-dose Chinese herbal medicine for warming kidney. The tamoxifen exerted no effects on the proliferation of normal human mammary epithelial cells in primary culture. CONCLUSION: The estradiol and progesterone can enhance the ability of proliferation of normal human mammary epithelial cells in primary culture. The regulating effect of Chinese herbal medicine for warming kidney on proliferation of normal human mammary epithelial cells in primary culture is dose-dependent.

[Discussion of relationship between quality of life and clinical effect assessment of malignant tumor treated with traditional Chinese medicine].

Traditional Chinese Medicine (TCM) therapy is effective in treating malignant tumor. TCM therapy is a multi-way, multi-layer and multi-target integrated treatment characterized by "survival with tumor" and improvement of life quality. An ideal result of the TCM therapy for cancer should be a positive response of the patient with satisfactory quality of life (QOL) and longer survival time. So it is necessary to introduce the QOL, quantification of symptoms and signs and standardization of TCM syndromes to the objective, quantitative and standardized effect assessment of TCM therapy for cancer. A criterion for evaluating the effect of TCM therapy on tumors should be established, which reflects the characteristics and advantages of the TCM therapy, and meets the requirement of the developing effect assessment of modern medicine in cancer treatment. It is suggested that the treatment effect should be the sum of quality adjusted life years (QALYs) (equal to QOL multiplied by time of survival), syndrome remission rate and tumor remission rate. But the proportion of each of the three is variable, which has close relationships with the different tumor stages and corresponding treatments.