Paraboeazunyiensis (Gesneriaceae), a new species from north Guizhou, China.

A new lithophytic species, Paraboeazunyiensis T.Deng, F.Wen & R.B.Zhang (Gesneriaceae), inhabiting Karst rocks in northern Guizhou, China, is introduced and depicted in this study. It bears a resemblance to P.crassifolia (Hemsl.) B.L. Burtt, yet is distinguishable by its shorter filaments and staminodes, triangular ovate calyx segments, and ovaries surpassing the styles in length. Moreover, the phylogenetic tree constructed from nuclear DNA (ITS) and plastid DNA (trnL-F) data firmly support the differentiation of this novel species from P.crassifolia.

Petrocodonwui (Gesneriaceae), a new species from Guizhou, China.

Petrocodonwui F.Wen & R.B.Zhang (Gesneriaceae), a typically lithophyte occurring in the Danxia areas of north-western Guizhou, China, is described and illustrated as new to science. The new species shows overall similarity with P.chishuiensis Z.B.Xin, F.Wen & S.B.Zhou, which is also its sister species, based on molecular evidence. The new species can be distinguished from P.chishuiensis by the elongated rhizome, the relatively long indumentum on the peduncle, the shape, size and indumentum of calyx lobes, the location of the stamens in the corolla tube and the shape, size and indumentum of the stigma. We provide a diagnosis, detailed description, photographic images and a table with taxonomic notes to distinguish several other morphologically similar Petrocodon species.

Long noncoding RNA Gm2694 drives depressive-like behaviors in male mice by interacting with GRP78 to disrupt endoplasmic reticulum homeostasis.

Long noncoding RNAs (lncRNAs) are involved in various biological processes and implicated in the regulation of neuronal activity, but the potential role of lncRNAs in depression remains largely unknown. Here, we identified that lncRNA Gm2694 was increased in the medial prefrontal cortex (mPFC) of male mice subjected to chronic social defeat stress (CSDS). The down-regulation of Gm2694 in the mPFC alleviated CSDS-induced depressive-like behaviors through enhanced excitatory synaptic transmission. Furthermore, we found that Gm2694 preferentially interacted with the carboxyl-terminal domain of 78-kilodalton glucose-regulated protein (GRP78), which abrogated GRP78 function and disrupted endoplasmic reticulum homeostasis, resulting in a reduction of the surface expression of AMPA receptors (AMPARs). Overexpression of GRP78 in the mPFC promoted the surface expression of AMPARs and attenuated the CSDS-induced depressive-like behaviors of mice. Together, our results unraveled a previously unknown role of Gm2694 in regulating endoplasmic reticulum homeostasis and excitatory synaptic transmission in depression.

Knockdown of POLQ interferes the development and progression of hepatocellular carcinoma through regulating cell proliferation, apoptosis and migration.

BACKGROUND: DNA Polymerase Theta (POLQ) is a DNA polymerase involved in error-prone translesion DNA synthesis (TLS) and error-prone repair of DNA double-strand breaks (DSBs), whose function in hepatocellular carcinoma has not been investigated. METHODS: In the present study, both the data collected from the Cancer Genome Atlas (TCGA) and our group's results showed higher POLQ expression in HCC tissues than the para-cancerous tissues, which was associated with higher malignancy and poor prognosis. POLQ knockdown HCC cell model (shPOLQ) was constructed along with the corresponding negative control (shCtrl) through lentivirus infection for loss-of-function study. RESULTS: We found that, upon knockdown of POLQ, the proliferation and migration of HCC cells decreased and apoptosis percentage increased. Moreover, the percentage of cells in G2 phase significantly increased in shPOLQ group compared with shCtrl group. Xenografts in mice grafted with shPOLQ cells grew much slower than that transplanted with shCtrl cells, and expressed lower Ki67 level. Furthermore, an apoptosis-related signaling array was used to explore the involvement of downstream signaling pathways, suggesting the enhanced phosphorylation of HSP27 and JNK, and the de-activation of mTOR, PRAS40, ERK1/2 and STAT3 pathways. CONCLUSIONS: Collectively, our study revealed that POLQ may participate in the development of HCC, depletion of which may be a promising treatment strategy for HCC.

[Short-term effect of intensity-modulated radiotherapy for children with high-risk neuroblastoma: an analysis of 24 cases].

OBJECTIVE: To study the efficacy and safety of intensity-modulated radiotherapy (IMRT) in children with high-risk neuroblastoma (NB). METHODS: A retrospective analysis was performed on the medical data of 24 children with high-risk NB who were diagnosed and treated with IMRT in the Department of Hematology and Oncology, Hunan Provincial People's Hospital, from April 2018 to December 2020. The medical data included age, radiotherapy dose, times of radiotherapy, laboratory examination results, adverse reactions, and survival. RESULTS: All 24 children (14 boys and 10 girls) received IMRT, with a mean age of (65±23) months and a median age of 59 months. The primary tumor was located in the abdomen in 23 children and 1 child had primary tumor in the mediastinum. The median age was 41.5 months at the time of radiotherapy. The radiation dose of radiotherapy ranged from 14.4 to 36.0 Gy, with a mean dose of (22±3) Gy and a daily dose of 1.8-2.0 Gy. The radiotherapy was performed for a total number of 8-20 times, with a mean number of 11.9 times. Among these children, 6 received radiotherapy for the residual or metastatic lesion. Of all the 23 children, 3 experienced cough, 2 experienced diarrhea, and 1 experienced vomiting during radiotherapy. At 2 weeks after radiotherapy, serum creatinine ranged from 2.3 to 70.1 µmol/L and alanine aminotransferase ranged from 9.1 to 65.3 µ/L. Ten children experienced grade Ⅲ bone marrow suppression and 2 experienced grade Ⅳ bone marrow suppression 1 to 2 weeks after radiotherapy. Four children experienced grade Ⅲ bone marrow suppression and 1 experienced grade Ⅳ bone marrow suppression 3 to 4 weeks after radiotherapy. During a median follow-up time of 13.5 months, 23 children (96%) achieved stable disease and 1 died. Up to the follow-up date, second malignant tumor or abnormal organ function was not observed. CONCLUSIONS: IMRT can improve the local control rate of NB. IMRT appears to be safe in the treatment of children with NB.

The retention characteristics for water-soluble and water-insoluble particulate matter of five tree species along an air pollution gradient in Beijing, China.

The air purification potential of plants has been widely studied and recognized. However, their specific capacities in retaining water-soluble (WSPM) and water-insoluble (WIPM) atmospheric particulate matter (PM) are still unclear. In order to recommend tree species with high air phytoremediation ability, the retention characteristics for WSPM and WIPM of five tree species under different haze pollution levels and PM retention durations in Beijing were evaluated after introducing ultrasonic cleaning procedure to the conventional leaf cleaning methods. The daily PM amount retained these species in the six central districts in Beijing (SCBD) was roughly estimated based on the field tree survey data in 171 plots randomly distributed within the Fifth Ring Road. The updated leaf cleaning method improved the evaluation accuracy for WSPM and WIPM by 54% and 31%, respectively. The particles retained by the broadleaf and coniferous species were mainly composed of WSPM (71%) and WIPM (64%), respectively. The diameter distribution of PM varied markedly with species, PM retention duration, and pollution level. However, it always showed a unimodal pattern for WSPM and no uniform patterns for WIPM. The average relative capacities of different species in retaining WSPM of TSP (PM ≤ 100 µm) were more stable with time, and the corresponding rank was Sophora japonica > Salix babylonica > Ginkgo biloba > Pinus tabuliformis > Sabina chinensis. Whereas, as to the WIPM of TSP, their order changed to S. japonica > P. tabuliformis > S. babylonica > G. biloba > S. chinensis. During the study period, the TPM (WIPM+WSPM) of TSP retained by these species per day in the SCBD reached 132.6 t (76.1 t WSPM + 56.5 t WIPM), accounting for a considerable proportion of the daily dust-fall amount. These findings can contribute to selecting greening tree species and managing the urban forest to improve urban air quality.

Paraphlomis kuankuoshuiensis (Lamiaceae), a new species from the limestone areas of northern Guizhou, China.

Paraphlomis kuankuoshuiensis (Lamiaceae), a new species found in the limestone areas of northern Guizhou, China, is described and illustrated in this paper. Based on its tubular-campanulate calyx, this taxon should be a member of sect. Paraphlomis Prain. The new species resembles P. patentisetulosa C.Y. Wu & H. W. Li, P. hispida C.Y. Wu, and P. hirsutissima C.Y. Wu & H.W. Li, but differs from these three taxa in the following aspects: the stems are very short (<7 cm), with one or two short internodes, giving the impression of having a tuft of basal leaves; it has sparsely setose hairs on the outer surface of the calyces and short fruiting calyces. The florescence, fruit period, habitat, and the geographical distribution of P. kuankuoshuiensis are also quite different from the three closely related species.

Effect of Bushen Tongluo Formula on Osteoporosis of Zebrafish and Study on Autophagy Mechanism of Osteoclast / 中国实验方剂学杂志

Objective::To evaluate the intervention effect of Bushen Tongluo formula on zebrafish osteoporosis model and to clarify its regulation of autophagy mechanism on osteoclast differentiation. Method::The 260 young zebrafishes (3 dpf) were selected, the zebrafish osteoporosis model was established with 25 mmol·L-1 prednisolone (Pred) for 3 days, confirming the successful model by calcein staining. The zebrafishes were divided into control group, Pred group (25 mmol·L-1), etidronate disodium (ED) group (300 mg·L-1), Bushen (BS) group (180 mg·L-1), Tongluo (TL) group (30 mg·L-1), and Bushen Tongluo (BSTL) group (210 mg·L-1), 40 tails per group. After intervened with medicine for 4 days, the calcein staining was adopted to count the vertebral bone fluorescence area of zebrafish, Real-time PCR was adopted to detect the mRNA expression of akaline phosphatase (ALP), bone morphogenetic protein 2b (BMP-2b), runt-related protein 2 (Runx2) and cathepsin K (CTSK), phosphorus and tartrate-resistant acid phosphatase (TRAP), nuclear factor of activated T-cells, cytoplasmic-1 (NFATC-1). Divided RAW264.7 cells into control, Rankl induction (10 μg·L-1), BS (180 mg·L-1), TL (30 mg·L-1), and BSTL group (210 mg·L-1) after they were cultured to 80%-90% density. The expression of actin ring was detected by phalloidin cytoskeleton staining. The mRNA expression of TRAP, CTSK and autophagy-related genes 5 (ATG5), autophagy-related genes 7 (ATG7), and ubiquitin-binding protein p62 (p62) were detected by Real-time PCR. Result::Compared with the control group in vivo, the vertebral area and ALP, BMP-2b, and Runx2 expression of zebrafish in the Pred group were significantly decreased (P<0.01), and CTSK, TRAP, and NFATC-1 expression of zebrafish were significantly increased (P<0.01). Compared with the Pred group, the vertebral area of the TL (P<0.05) and BSTL group (P<0.01) increased significantly. The expressions of ALP, BMP-2b, and Runx2 in the BS, TL, and BSTL group were significant increased (P<0.01). The expression of CTSK, TRAP, and NFATC-1 in BSTL group were significant decreased (P<0.05, P<0.01). Compared with the control group in vitro, the number of actin rings (P<0.01) and CTSK, TRAP, ATG5 (P<0.01) expression and ATG7, p62 (P<0.01) expression were significantly increased in the Rankl-induced group. Compared with the Rankl induction group, the number of actin rings and CTSK and TRAP expression were significantly decreased in the BS, TL, and BSTL group (P<0.05, P<0.01), while the expression of ATG5, ATG7, and p62 were significant decreased in the BSTL group (P<0.05). Conclusion::BSTL can significantly improve prednisolone-induced zebrafish osteoporosis and inhibit osteoclast differentiation by reducing autophagy-related gene expression.

Sedum lipingense (Crassulaceae) identifying a new stonecrop species in SE Guizhou, China, based on morphological and molecular evidence.

We describe and illustrate Sedum lipingense (Crassulaceae), a new species of stonecrop found in the limestone areas of SE Guizhou, China. Based on the presence of adaxially gibbous carpels and follicles, this taxon belongs to sect. Sedum S.H. Fu. The new species superficially resembles S. subtile Miquel and S. bulbiferum Makino but differs from these two taxa in its development of a basal leaf rosette during florescence. The nrDNA internal transcribed spacer (ITS) sequences also support the claim that this plant is a new species in the Sedum genus.

Primulina serrulata (Gesneriaceae), a new species from southeastern Guizhou, China.

Primulina serrulata R.B.Zhang & F. Wen, a new species from a limestone area in southeastern Guizhou, China, is described and illustrated here. The new species is morphologically related to P. fimbrisepala (Hand.-Mazz.) Y.Z.Wang. We examined the morphological differences between these congeners and provide illustrations and photographs of this new species in this paper.

Effective targeted therapy based on dynamic monitoring of gene mutations in non-small cell lung cancer.

With the rapid development of precision medicine, next generation sequencing (NGS) has provided the ability to decode tumors at the DNA level. In treatment of patients with non-small cell lung cancer (NSCLC), it is of great importance to identify epidermal growth factor receptor (EGFR) mutations and drug resistance mechanisms in the late stages. A Chinese Han male patient (64 years old) who was initially diagnosed with EGFR-19-deletion-positive advanced NSCLC had a satisfactory clinical response after treatment with erlotinib. Subsequently, the disease progressed and NGS in plasma-derived circulating tumor DNA (ctDNA) revealed T790M mutation. The patient was then treated with osimertinib. In addition, NGS in ctDNA was performed again after the disease progressed, suggesting that MET was amplified, and then the patient was alternatively treated with combination therapy of crizotinib and erlotinib. The disease progressed for the third time, and treatment with osimertinib was undertaken again according to the T790M testing results. Dynamic monitoring of ctDNA was found to be helpful in selecting the appropriate treatment methods and prolonging the survival time of the patient.

Calycosin Inhibits Intestinal Fibrosis on CCD-18Co Cells via Modulating Transforming Growth Factor-ß/Smad Signaling Pathway.

BACKGROUND: Intestinal fibrosis is the major complication of Crohn's disease (CD). There are no other good treatments for CD except surgery and remains a refractory disease. Calycosin (CA), the active component of astragalus membranaceus, has been reported the potential effect on lung fibrosis and renal fibrosis. In this study, we aim to explore the effect of CA on intestinal fibrosis in vitro and the possible signal pathway. METHODS: The antifibrotic effect of CA is investigated in human intestinal fibroblasts (CCD-18Co) cells induced by transforming growth factor-ß1 (TGF-ß1). MTT method was used to screen the concentration of CA. Real-time polymerase chain reaction and western blot analysis were used to evaluate the expression of α-smooth muscle actin (α-SMA), collagen I, and TGF-ß/Smad pathway. RESULTS: The results showed that the concentration of CA was 12.5, 25, 50 µmol/L. CA could inhibit the expression of α-SMA and collagen I. In addition, CA regulated the expression of TGF-ß/Smad signaling pathway. CONCLUSION: This study demonstrated that CA could inhibit the activation of CCD-18Co cells and reduce the expression of extracellular matrix. Our study highlighted that CA-inhibited TGF-ß/Smad pathway through inhibiting the expression of p-Smad2, p-Smad3, Smad4, and TGF-ß1 and raised the Smad7 expression. Therefore, CA might inhibit intestinal fibrosis by inhibiting the TGF-ß/Smad pathway.

Inhibition effect of phytoestrogen calycosin on TGF-ß1-induced hepatic stellate cell activation, proliferation, and migration via estrogen receptor ß.

The present study was designed to investigate the effects of calycosin on hepatic stellate cell (HSC) function and to explore whether the drug exerts its effect through the estrogen receptor. HSC proliferation and migration were measured by MTT assay and transwell chamber assay, respectively. The mRNA and protein expression of α-SMA, COL-I, and ERß were detected by real-time PCR and Western blotting. The co-localization and expression of α-SMA and ERß protein were detected by immunofluorescence. All the studies were investigated in the absence or presence of ICI 182,780. The results showed that calycosin inhibited the proliferation of activated HSCs and remarkably inhibited HSC migration. Calycosin significantly reduced the expression of α-SMA and COL-I in activated HSCs. However, with co-treatment with ICI 182,780, the inhibitory effect of calycosin against the above effects was strongly negated. Importantly, calycosin significantly downregulated the expression of ERß protein, while co-treatment with ICI 182,780 partially reversed the ERß downregulation. In addition, α-SMA decreased with the decrease of ERß expression and the subtype of ERß on HSC is ERß5. In conclusion, calycosin inhibits proliferation, activation, and migration of TGF-ß1-induced HSCs. The effect may be related to binding and downregulation of ERß5.

A low serum Tat-interacting protein 30 level is a diagnostic and prognostic biomarker for hepatocellular carcinoma.

The present study aimed to evaluate the diagnostic and prognostic value of Tat-interacting protein 30 (HTATIP2/TIP30) levels alone and in combination with α-fetoprotein (AFP) for the evaluation of hepatocellular carcinoma (HCC) patients. ELISA and immunohistochemical measurements on the serum and tissue of HTATIP2/TIP30 protein from HCC patients and normal controls were made. Receiver operating characteristic (ROC) curve analyses of AFP and HTATIP2/TIP30 were performed, as well as logistic regression analysis of APF combined with HTATIP2/TIP30. Log-rank analysis was used to correlate the prognosis with various levels of HTATIP2/TIP30. HTATIP2/TIP30 levels were significantly lower in the HCC group compared with the control group (4.50±2.63 vs. 9.50±2.04 ng/ml, P<0.001). ROC analysis revealed an optimal cut-off point at 7.27 ng/ml HTATIP2/TIP30 for separating the HCC from the control groups. The sensitivity and specificity were 84.6 and 93.7% (P<0.001), respectively. ROC areas of HTATIP2/TIP30 (0.928, P<0.001) were significantly higher than those for AFP (P<0.001). The area under the curve of the HTATIP2/TIP30 and AFP combination was 0.950 (P<0.001). Log-rank tests revealed that the recurrence-free survival time of the group with HTATIP2/TIP30>5.71 ng/ml was significantly higher than that of the control group (P<0.001). This is the first study to demonstrate that HTATIP2/TIP30 levels in serum may be an effective biomarker for the diagnosis and prognosis of HCC.

microRNA-141 is involved in a nasopharyngeal carcinoma-related genes network.

microRNAs (miRNAs) are small non-coding RNAs and have been implicated in the pathology of various diseases, including cancer. Here we report that the miRNA profiles have been changed after knockdown of one of the most important oncogene c-MYC or re-expression of a candidate tumor suppressor gene SPLUNC1 in nasopharyngeal carcinoma (NPC) cells. Both c-MYC knockdown and SPLUNC1 re-expression can down-regulate microRNA-141 (miR-141). miR-141 is up-regulated in NPC specimens in comparison with normal nasopharyngeal epithelium. Inhibition of miR-141 could affect cell cycle, apoptosis, cell growth, migration and invasion in NPC cells. We found that BRD3, UBAP1 and PTEN are potential targets of miR-141, which had been confirmed following luciferase reporter assays and western blotting. BRD3 and UBAP1 are both involved in NPC carcinogenesis as confirmed through our previous studies and PTEN is a crucial tumor suppressor in many tumor types. BRD3 is involved in the regulation of the Rb/E2F pathway. Inhibition of miR-141 could affect some important molecules in the Rb/E2F, JNK2 and AKT pathways. It is well known that carcinogenesis of NPC is involved in the networks of genetic and epigenetic alteration events. We propose that miR-141- and tumor-related genes c-MYC, SPLUNC1, BRD3, UBAP1 and PTEN may constitute a gene-miRNA network to contribute to NPC development.

Promoter methylation inhibits BRD7 expression in human nasopharyngeal carcinoma cells.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a head and neck malignancy with high occurrence in South-East Asia and Southern China. Recent findings suggest that epigenetic inactivation of multiple tumor suppressor genes plays an important role in the tumourigenesis of NPC. BRD7 is a NPC-associated bromodomain gene that exhibits a much higher-level of mRNA expression in normal than in NPC biopsies and cell lines. In this study, we explored the role of DNA methylation in regulation of BRD7 transcription. METHODS: The presence of CpG islands within BRD7 promoter was predicted by EMBOSS CpGplot and Softberry CpGFinder, respectively. Nested methylation-specific PCR and RT-PCR were employed to detect the methylation status of BRD7 promoter and the mRNA expression of BRD7 gene in tumor cell lines as well as clinical samples. Electrophoretic mobility shift assays (EMSA) and luciferase assay were used to detect the effects of cytosine methylation on the nuclear protein binding to BRD7 promoter. RESULTS: We found that DNA methylation suppresses BRD7 expression in NPC cells. In vitro DNA methylation in NPC cells silenced BRD7 promoter activity and inhibited the binding of the nuclear protein (possibly Sp1) to Sp1 binding sites in the BRD7 promoter. In contrast, inhibition of DNA methylation augments induction of endogenous BRD7 mRNA in NPC cells. We also found that methylation frequency of BRD7 promoter is much higher in the tumor and matched blood samples from NPC patients than in the blood samples from normal individuals. CONCLUSION: BRD7 promoter demethylation is a prerequisite for high level induction of BRD7 gene expression. DNA methylation of BRD7 promoter might serve as a diagnostic marker in NPC.

Effect of SPLUNC1 protein on the Pseudomonas aeruginosa and Epstein-Barr virus.

Short palate, lung and nasal epithelium clone 1 (SPLUNC1) gene coded a secreted protein found at the surface of nasopharyngeal epithelium, which may be an innate immunity defensive molecular and a risk factor for nasopharyngeal carcinoma (NPC). Here, we observed the effects of SPLUNC1 on the Gram negative bacteria Pseudomonas aeruginosa, evaluated the ability of SPLUNC1 protein binding to lipopolysaccharide. To observe the effect of SPLUNC1 protein on Epstein-Barr virus (EBV), we raised three EBV-transformed B-lymphocyte lines and treated the cells by SPLUNC1 protein; cellular disruption, apoptosis, EBV DNA content, and viral oncogene expression were analyzed. We found that SPLUNC1 protein can bind to bacterial lipopolysaccharide, inhibit the growth of P. aeruginosa, enhance the disruption and apoptosis of EBV-infected B-lymphocytes, downregulate protein expression of EBV latent membrane protein 1, while upregulate protein expression of EBV envelope glycoprotein gp350/220. The total EBV DNA in the culture medium was decreased significantly after 7 days of treatment by SPLUNC1. This study shows that SPLUNC1 not only has the role of antibacteria and antivirus, but also inhibits the potential oncogenicity of EBV in respiratory epithelium.

[Effect of growth inhibition of the secretory protein SPLUNC1 on Pseudomonas aeruginosa].

OBJECTIVE: To express the recombinant SPLUNC1 protein in HNE1 cells and to study its function of bactericidal and binding to lipopolysaccharide (LPS). METHODS: Full length of SPLUNC1 gene was cloned into pCMV-tag4A vector and stably transfected into HNE1 cell lines, the supernatant of cell cultures was collected. After being treated with the supernatant, the Pseudomonas aeruginosa was seeded to LB soft agar plate, and the bacteria clones were counted and analyzed. For in vitro LPS binding assay, LPS was coated to 96-well plates. We incubated in the plate with SPLUNC1 protein, and detected the binded SPLUNC1 protein by ELISA. Incubating the FITC-LPS with the SPLUNC1 stably transfected or control cells, the intracellular intensity of fluorescence was observed under the fluorescence microscope. RESULTS: SPLUNC1 inhibited the bacteria clone formation obviously. Although the binding efficiency of LPS and SPLUNC1 in vitro was very low, more FITC-LPS entered into the SPLUNC1 stably transfected cells. CONCLUSION: SPLUNC1 can inhibit the growth of Pseudomonas aeruginosa and bind LPS, and play an important defensive role in innate immunity of the upper airway.