Simultaneous Preparation of Abundant Flavonol Triglycosides from Tea Leaves.

Flavonol glycosides are important components of tea leaves, contributing to the bioactivities as well as bitterness and astringency of tea. However, the standards of many flavonol triglycosides are still not available, which restricts both sensory and bioactivity studies on flavonol glycosides. In the present study, we established a simultaneous preparation method of seven flavonol triglycoside individuals from tea leaves, which consisted of two steps: polyamide column enrichment and preparative HPLC isolation. The structures of seven flavonol triglycoside isolates were identified by mass and UV absorption spectra, four of which were further characterized by nuclear magnetic resonance spectra, namely, quercetin-3-O-glucosyl-rhamnosyl-glucoside, quercetin-3-O-rhamnosyl-rhamnosyl-glucoside, kaempferol-3-O-glucosyl-rhamnosyl-glucoside and kaempferol-O-rhamnosyl-rhamnosyl-glucoside. The purities of all isolated flavonol triglycosides were above 95% based on HPLC, and the production yield of total flavonol glycosides from dry tea was 0.487%. Our study provides a preparation method of flavonol triglycosides from tea leaves, with relatively low cost of time and solvent but high production yield.

Bursopentin (BP5) induces G1 phase cell cycle arrest and endoplasmic reticulum stress/mitochondria-mediated caspase-dependent apoptosis in human colon cancer HCT116 cells.

BACKGROUND: Bursopentin (BP5) is a multifunctional pentapeptide found in the chicken bursa of Fabricius. Recent study indicated that BP5 significantly stimulates expression of p53 protein in colon cancer HCT116 cells. However, the effects and underlying mechanisms of BP5 on HCT116 cell proliferation remain largely unclear. METHODS: Analyses of cell viability, cell cycle arrest as well as apoptosis were performed to study the actions of BP5 on HCT116 cells. Western blot analyse was assayed to measure the cell cycle-related and apoptosis-related proteins. Specific siRNAs targeting IRE1, ATF-6, and PERK were used for IRE1, ATF-6, and PERK knockdown, respectively. Cellular reactive oxygen species (ROS) were detected using a H2DCF-DA green fluorescence probe. Cytosolic free Ca2+ concentrations and mitochondrial membrane potential (ΔΨm) were measured using Fluo-3 AM and JC-1 stains, respectively. RESULTS: BP5 possessed strong inhibitory effects on the cell growth and induced apoptosis in HCT116 cells. Mechanistically, BP5 arrested the cell cycle at G1 phase by increasing p53 and p21 expression and decreasing cyclin E1-CDK2 complex expression. BP5 treatment dramatically activated the endoplasmic reticulum (ER) stress-mediated apoptotic pathway, as revealed by the significantly enhanced expression of unfolded protein response (UPR) sensors (IRE1α, ATF6, PERK) as well as downstream signaling molecules (XBP-1s, eIF2α, ATF4 and CHOP), and by the significantly altered the BP5-induced phenotypic changes in IRE1, ATF6, and PERK knockdown cells. Additionally, BP5-induced ER stress was accompanied by the accumulation of cytosolic free Ca2+ and intracellular ROS. Furthermore, BP5 treatment resulted in the increase of Bax expression, the decrease of Bcl-2 expression and the reduction of ΔΨm, subsequently causing a release of cytochrome c from the mitochondria into the cytoplasm and finally enhancing the activities of caspase-9 and -3. In addition, z-VAD-fmk, a pan-caspase inhibitor, markedly rescued BP5-induced cell viability reduction and reduced BP5-induced apoptosis. CONCLUSIONS: Our present results suggest that BP5 has an anticancer capacity to arrest cell cycle at G1 phase and to trigger ER stress/mitochondria-mediated caspase-dependent apoptosis in HCT116 cells. Therefore, our findings provide insight into further investigations of the anticancer activities of BP5.

Overexpression of MTHFD1 in hepatocellular carcinoma predicts poorer survival and recurrence.

Aim: MTHFD1 was the enzyme providing one-carbon derivatives of tetrahydrofolate. We sought to investigate the impact of MTHFD1 on hepatocellular carcinoma (HCC). Methods: Bioinformatic analysis, western blot and immunohistochemistry were conducted to detect MTHFD1 expression in HCC. The relationships between MTHFD1 and prognosis of 172 HCCs were analyzed by Kaplan-Meier method and Cox proportional hazards model. Results: High MTHFD1 expression in HCC represented poor prognosis (overall survival p = 0.025; time to recurrence p = 0.044). Combining MTHFD1 with serum AFP, survival analysis demonstrated the prognosis of the MTHFD1 low expression and AFP ≤20 ng/ml group was better than that of the MTHFD1 high expression or AFP >20 ng/ml group and the MTHFD1 high expression and AFP >20 ng/ml group (overall survival p < 0.0001; time to recurrence p < 0.0001). Conclusion: High MTHFD1 expression in HCC indicated poorer prognosis. Combining MTHFD1 with serum AFP improved the accuracy of prognostic prediction.

A new simple index for characterizing the labile heavy metal concentration in soil by diffusive gradients in thin films technique.

Diffusive gradients in thin films technique (DGT) is recognized as a more reliable method for determining labile heavy metal (HM) concentration in soil than traditional destructive methods. However, the current DGT measurement index, CDGT, theoretically underestimates the true labile concentration (Clabile) of HMs in soil and lacks direct comparability with the conventional soil HM content indices due to unit differences. Here, we proposed CDGT-W, a new simple index which is defined as the HM accumulation in the binding layer, normalized to the weight of soil (optimized water content = 100% of the maximum water holding capacity) filled in the open cavity-type DGT device over a specified deployment time (optimized time = 24 h). The procedure for measuring CDGT-W is analogous to that of CDGT but includes precise determination of water content (water/dry soil) and the mass of soil filled in the cavity. We conducted measurements of Cu, Pb, Cr(Ⅵ) and As(V) as CDGT-W, CDGT, solution concentration (Csoln), and CaCl2 extractable concentration (CCaCl2) on three soils with a diverse range of HM concentrations. CDGT-W showed significant linear correlations with all other tested indexes. The ratios of CDGT-W to CCaCl2 varied between 0.30 and 0.98 for all HM-soil combinations with only one exception, a range much greater than CDGT/Csoln (typically <0.1) but lower than 1. This suggested that CDGT-W may more accurately reflect Clabile than CDGT (theoretically underestimates Cliable) and CCaCl2(likely overestimates Cliable). Additionally, CDGT-W measurements for these four HMs exhibited a broad measure concentration range and a low detection limit (mg/kg level). Consequently, CDGT-W may offer a more reliable alternative to CDGT for characterizing Clabile in unsaturated soils.

Structural characterization and immune activity evaluation of a polysaccharide from Lyophyllum Decastes.

An innovative acidic hydrolysate fingerprinting workflow was proposed for the characterization of Lyophyllum Decastes polysaccharide (LDP) by ultra performance liquid chromatography-mass spectrometry (UPLC-MS). The crude polysaccharides were firstly separated and purified by using DE-52 column and the BRT GPC purification system, respectively. The molecular weight and monosaccharide content of homogeneous polysaccharides were ascertained by utilizing HPGPC and ion chromatography separately. Secondly, the linkage of LDP was identified by methylation analysis and 1D/2D NMR spectra. The UPLC-MS/MS was used to scan and identify the acidic hydrolysate products of LDP using the PGC column. The oligosaccharides were collected by chromatography and identified by mass spectrometry. Thirdly, the expression of IL-1ß, IL-6, iNOS, TNF-α and IFNAR-I was measured in order to assess the immunological activity of LDP. Besides, the targeted receptors identification of polysaccharides was performed by screening the expression of TLRs family protein. The results showed that oligosaccharide fragments with different molecular weights can be obtained by partial hydrolysis, which further verified that the structures of LDP polysaccharides was a 1-6-linked ß-glucan. Moreover, the LDP polysaccharide can up-regulate the content of IL-1ß, IL-6, iNOS, TNF-α and IFNAR-I and plays an important immunoregulation role through TLRs family.

Inulin-type fructans obtained from Atractylodis Macrocephalae by water/alkali extraction and immunoregulatory evaluation.

Two homogenous polysaccharides extracted from Atractylodes macrocephala Koidz. were investigated by water extraction (AMP-FW) and alkali solution extraction (AMP-FA) after purification by anion exchange column and size exclusion chromatography. The molecular weight of AMP-FW and AMP-FA were 2874 Da and 3438 Da, respectively, estimated by high performance gel permeation chromatography (HPGPC). The monosaccharide compositions of AMP-FW and AMP-FA were glucose and fructose at a molar ratio of 0.11:0.89 determined by high performance anion exchange chromatography (HPAEC). The functional groups, glycosidic linkages and the chemical structure were characterized by FT-IR, GC-MS and NMR, which comprehensively indicated a similar inulin-type fructan structure of the two polysaccharides from A. macrocephala. However, the scanning electron microscopy (SEM) results showed different microstructures that irregular lamellar shape for the AMP-FW and spheroid shape for the AMP-FA. The further studies on immunomodulation showed that AMP-FW at 50 µg/mL could significantly (P < 0.05) stimulate RAW 264.7 cells by enhancing the mRNA expression of TNF-α and IL-1ß, which had a relative high immunomodulatory potential when compared to AMP-FA. Their activation on different toll-like receptors (TLR) also indicated their different roles in the immunoregulation. Overall, these findings reported here will serve as the basis for further structure-activity relationship studies.

[Inhibitory effect of 5-fluorouracil on HT-1080 human fibrosarcoma cells in vitro].

OBJECTIVE: To evaluate the inhibitory effect of 5-fluorouracil (5-Fu) on HT-1080 human fibrosarcoma cells in vitro. METHODS: HT-1080 human fibrosarcoma cells were cultured for 3 days in the proliferation period. Then adriamycin or 5-Fu at different concentrations were used to treat these cells for 24 h, 72 h and 144 h. MTT assay was used to evaluate the cytotoxic effects through measuring optical density and calculating the inhibition rate of cell growth. Morphologic changes of the cells were observed under a phase contrast microscope. Flow cytometry (FCM) was performed to detect the changes in cell cycle and DNA ploidy in the fibrosarcoma cells treated with 5-Fu. RESULTS: 10 µg/ml 5-Fu showed an inhibition rate of 45.9% (24 h), 64.7% (72 h) and 90.6% (144 h) of the HT-1080 cell growth. 100 µg/ml 5-Fu showed an inhibition rate of 53.1% (24 h), 86.4% (72 h), 93.0% (144 h) of the HT-1080 cell growth, results similar to those in the adriamycin group. Untreated fibrosarcoma cells accounted for 67.5% in G(1) phase, 21.2% in S phase and 11.3% in G(2) phase. With the increasing drug concentrations, cells in G(1) + S phase increased rapidly and no cells in G(2) phase were observed later. The cells treated with 5-Fu showed a G(1) + S cell cycle arrest. CONCLUSIONS: 5-Fu has an antitumor activity in human fibrosarcoma HT-1080 cells in vitro, in a time-dependent and dose-dependent manner. The cytotoxity of 5-Fu at high concentrations and continuous use can induce tumor cell cycle arrested at G(1) + S phase, a similar result induced with adriamycin.

Expression and prognosis analysis of PAQR5 in kidney cancer.

Progestin and adipoQ receptor 5 (PAQR5) affects the development of various malignancies and is specifically expressed in kidney. However, the role of PAQR5 in renal carcinoma remains unclear. We assessed the state of PAQR5 expression in kidney renal clear cell carcinoma (KIRC) by The Cancer Genome Atlas and Gene Expression Omnibus datasets. Moreover, immunohistochemistry was performed to observe the expressions of PAQR5 protein in tumor tissues. The relationships between PAQR5 expression and clinical characteristics were investigated by UALCAN. Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan-Meier plotter were used to analyze the effect of PAQR5 expression levels on overall survival and relapse-free survival (RFS). The re lationships between clinical characteristics and survival were also evaluated by univariate and multifactorial Cox regression. Gene Ontology term analysis, Kyoto Encyclopedia of Genes and Genomes analysis, and gene set enrichment analysis were performed on PAQR5 to explain the enrichment pathways and functions. Protein and protein interactions were explained by GeneMANIA and STRING. We also explored the relevance of PAQR5 to tumor immune cell infiltration and immunomodulatory molecules by TIMER and GEPIA. Finally, we explored the correlation of PAQR5 with the pathway proteins STATs, HIF-1α, and mTOR using the GSE40435 dataset. PAQR5 expression was low in KIRC and correlated significantly with clinical characteristics including cancer stage, tumor grade, and nodal metastasis status. Low PAQR5 expression was significantly associated with poorer survival. Cox regression analysis indicated that upregulation of PAQR5 was an independent factor for a good prognosis of KIRC. PAQR5 downregulation was associated mainly with STAT3 target upregulation, tumorigenesis, and poor differentiation. PAQR5 expression also correlated positively with B cells, neutrophils, macrophages, and dendritic cells and negatively with the infiltration of FOXP3+ Treg cells and the immune checkpoint molecules PD-1, CTLA4, and LAG3. Moreover, PAQR5 expression in KIRC was negatively correlated with the pathway proteins STAT1/2/3/4/5A, HIF-1α, and mTOR. PAQR5 is an excellent predictor of KIRC prognosis and may be a potential molecular therapeutic target.

Phase I study of pegylated liposomal doxorubicin and cisplatin in patients with advanced osteosarcoma.

PURPOSE: The repeated use of doxorubicin is limited due to dose-limiting cardiac toxicity. Pegylated liposomal doxorubicin (PEG-LD, Duomeisu) has a reduced cardiac toxicity. This phase I study aimed to investigate the maximum tolerated doses (MTDs) and dose-limiting toxicities (DLTs) of the PEG-LD and cisplatin combination in patients with metastatic and recurrent osteosarcoma. METHODS: Patients were given PEG-LD at a dose of 40, 50, or 60 mg/m2 on day 1 of each 21-day cycle, according to a 3 + 3 approach for dose escalation. Cisplatin was administered as a fixed dose of 100 mg/m2 for every cycle. Toxicities and tumor response were observed. RESULTS: A total of 15 patients were enrolled in this trial, and nine of the patients had received prior doxorubicin. The MTD of PEG-LD was reached at 50 mg/m2 in this regimen, with neutropenic fever and stomatitis as DTLs. The main adverse event (AE) was myelosuppression. The most common non-hematological AEs were vomiting, hypoproteinemia, stomatitis and transient sinus arrhythmia. Grade 3-4 toxicity was neutropenia, leukopenia, thrombocytopenia, anemia and stomatitis in the whole cohort. All the AEs were relieved after symptomatic and supportive treatment. Totally, the overall response rate was 13.3% and disease control rate was 66.7%. For the six patients who have not received prior doxorubicin, one partial response and five stable diseases were observed. CONCLUSION: We provide the data showing that PEG-LD 50 mg/m2 combined with cisplatin 100 mg/m2 demonstrated an acceptable safety profile and promising clinical activity in advanced osteosarcoma, which merits further evaluation in phase II studies. TRIAL REGISTRATION: ChiCTR1900021550.

[Retrospective analysis and prognostic factors of 208 cases of primary soft tissue sarcoma of extremity].

OBJECTIVE: To investigate the prognosis factors of soft tissue sarcoma, especially the impact of surgical treatment on the prognosis. METHODS: We retrospectively reviewed 208 surgically treated patients. There were 128 male and 80 female. The average age was 46 ranged from 9 to 98 years old. Possible factors of whether the patient firstly treated in our hospital, the tumor size (< 5 cm, 5 ∼ 10 cm, > 10 cm), tumor depth (superficial deep fascia, under the deep fascia), histological type (such as adipose sarcoma, malignant fibrous histiocytoma, synovial sarcoma, fibrous sarcoma, malignant peripheral nerve sheath tumors, other tumors), tumor grade (FNCLCC I, II, III), surgical margin (intralesional, marginal, wide, radical) and adjuvant therapy on the prognosis of patients were analyzed. RESULTS: The median follow-up was 37.5 ranged from 1.3 to 128.1 months. The overall 3-year and 5-year survival were 77% and 75%. The overall 3-year and 5-year recurrence rate were 28% and 37%. The overall 3-year and 5-year metastasis rate were 35% and 43%. Tumor size, tumor grade and metastasis or not independently affected survival (χ(2) = 18.813, 24.849 and 21.107, all P < 0.05). Whether the patient firstly treated in our hospital and histological type independently affect the local recurrence (χ(2) = 21.915, 12.192, both P < 0.05); histological grade can independently affect the metastasis (χ(2) = 7.714, P < 0.05). Surgical margin alone affected the local recurrence and metastasis (χ(2) = 19.610, 9.272, both P < 0.05). CONCLUSIONS: Surgical margin independently affected local recurrence and distant metastasis, and thus indirectly affect the survival of soft tissue sarcoma. In particular, the primary choice for treatment of soft tissue sarcoma without metastasis should be surgery. Wide or radical margin could significantly improve the prognosis of soft tissue sarcoma patients.

Frequent amplification of HDAC genes and efficacy of HDAC inhibitor chidamide and PD-1 blockade combination in soft tissue sarcoma.

BACKGROUND: The advent of immune checkpoint therapy has been a tremendous advance in cancer treatment. However, the responses are still insufficient in patients with soft tissue sarcoma (STS). We aimed to identify rational combinations to increase the response to immune checkpoint therapy and improve survival. METHODS: Whole-exome sequencing (WES) was performed in 11 patients with liposarcoma. Somatic copy number alterations (SCNAs) were analyzed at the gene level to identify obvious amplification patterns in drug-target genes. The expression and prognostic value of class I histone deacetylases (HDACs) was evaluated in 49 patients with sarcoma in our center and confirmed in 263 sarcoma samples from The Tumor Cancer Genome Atlas (TCGA) database. Q-PCR, flow cytometry and RNA-seq were performed to determine the correlations between class I HDACs, chidamide and PD-L1 in vitro and in vivo. The efficacy of combining chidamide with PD-1 blockade was explored in an immunocompetent murine model and a small cohort of patients with advanced sarcoma. Western blot, ChIP assay and dual luciferase assessment were applied in the mechanistic study. RESULTS: The HDAC gene family was frequently amplified in STS. SCNAs in the HDAC gene family were extensively amplified in 8 of 11 (73%) patients with liposarcoma, based on a drug-target gene set, and we verified amplification in 76.65% (197/257) of cases by analyzing TCGA sarcoma cohort. Class I HDAC expression is associated with a poor prognosis for patients with STS, and its inhibition is responsible for promoting apoptosis and upregulating of programmed cell death ligand 1 (PD-L1). The HDAC class I inhibitor chidamide significantly increases PD-L1 expression, increased the infiltration of CD8+ T cells and reduced the number of MDSCs in the tumor microenvironment. The combination of chidamide with an anti-PD-1 antibody significantly promotes tumor regression and improves survival in a murine model. Moreover, chidamide combined with the anti-PD-1 antibody toripalimab is effective in patients with advanced and metastatic sarcoma, and the side effects are tolerable. Mechanistically, chidamide increases histone acetylation at the PD-L1 gene through the activation of the transcriptional factor STAT1. CONCLUSIONS: The combination of chidamide and anti-programmed cell death 1 (PD-1) therapy represents a potentially important strategy for STS.

Clinical observations on 46 cases of globus hystericus treated with modified Banxia Houpu decoction.

OBJECTIVE: To observe the therapeutic effect and mechanism of modified Banxia Houpu decoction on globus hystericus. METHODS: The 95 patients with globus hystericus were randomly divided into a treatment group of 46 cases treated with modified Banxia Houpu decoction and a control group of 49 cases treated with Manyanshuning (Granula for Clearing the Throat). In addition, a normal group of 24 healthy people was set up. SCL-90 scale was adopted to observe the therapeutic effect, evaluate the psychological state of patients and build a database on combination of four diagnoses. RESULTS: The effect of the modified Banxia Houpo decoction was better than that of the control group in relieving depression, anxiety and improving the psychological state (P<0.05 or P<0.01). CONCLUSION: Modified Banxia Houpu decoction has definite therapeutic effect on globus hystericus. Its mechanism may be related to its function in relieving depression and anxiety and regulating the psychological state.

[Surgical treatment for bone nonunion after massive allograft transplantation].

OBJECTIVE: To evaluate the surgical treatment and outcome of autogenous bone grafting and internal fixation in management of bone nonunion after massive allograft transplantation. METHODS: From January 1994 to December 2006, 41 of 176 patients underwent bone nonunion after massive allograft transplantation. Twenty-two of 41 patients received autogenous bone grafting. Complete clinical and follow-up data was available for 15 cases. The average age at secondary autogenous bone grafting was 24 years old (ranging from 15 to 34). The primary diseases included osteosarcoma (5 cases), giant cell tumor (4 cases), parosteal osteosarcoma (2 cases), hemangioendothelioma (2 cases) and primitive neuroectodermal tumor (2 cases). Tumor was located at distal femur in 7 patients, middle of humerus in 3, middle of femur in 2, proximal tibia in 2 and proximal humerus in 1. Eight of 15 patients with simple bone nonunion received autogenous bone grafting. Another 7 patients with bone nonunion and fracture of primary internal fixation underwent autogenous bone grafting and re-internal fixation. RESULTS: At a mean follow-up of 46.8 months (ranging from 18 to 148 months), bone union was observed in 13 of 15 patients (86.7%) with the mean healing time 13.3 months (ranging from 5 to 20). Bone union could be observed in all 8 patients with simple bone nonunion and 5 of 7 patients with bone nonunion and internal fixation fracture, similar healing time 14 and 12 months respectively. There was no infection or any other complications. Two patients underwent re-nonunion received prosthesis replacement at last. The mean MSTS score of 13 patients was 25.1, with 8 simple bone nonunion patients and 5 combined with internal fixation fracture patients 25.4 and 24.6 respectively, also basically no difference. CONCLUSIONS: Autogenous bone grafting and internal fixation in management of nonunion after massive allograft transplantation have the advantage of easy operation, less complications, high rate of bone healing and good function result with obvious superiority to prosthesis replacement. For management of nonunion after massive allograft transplantation, autogenous bone grafting and internal fixation is mostly recommended.

[The status and prospect of total body irradiation technique].

This paper describes the development of TBI technique about fractionated TBI and the relationship between irradiation dose rate and complication; IMRT for TBI and lung compensation technique.

Amnioinfusion for the prevention of the meconium aspiration syndrome.

BACKGROUND: It is uncertain whether amnioinfusion (infusion of saline into the amniotic cavity) in women who have thick meconium staining of the amniotic fluid reduces the risk of perinatal death, moderate or severe meconium aspiration syndrome, or both. METHODS: We performed a multicenter trial in which 1998 pregnant women in labor at 36 or more weeks of gestation who had thick meconium staining of the amniotic fluid were stratified according to the presence or absence of variable decelerations in fetal heart rate and then randomly assigned to amnioinfusion or to standard care. The composite primary outcome measure was perinatal death, moderate or severe meconium aspiration syndrome, or both. RESULTS: Perinatal death, moderate or severe meconium aspiration syndrome, or both occurred in 44 infants (4.5 percent) of women in the amnioinfusion group and 35 infants (3.5 percent) of women in the control group (relative risk, 1.26; 95 percent confidence interval, 0.82 to 1.95). Five perinatal deaths occurred in the amnioinfusion group and five in the control group. The rate of cesarean delivery was 31.8 percent in the amnioinfusion group and 29.0 percent in the control group (relative risk, 1.10; 95 percent confidence interval, 0.96 to 1.25). CONCLUSIONS: For women in labor who have thick meconium staining of the amniotic fluid, amnioinfusion did not reduce the risk of moderate or severe meconium aspiration syndrome, perinatal death, or other major maternal or neonatal disorders.

[Design of a 3D afterloading brachytherapy simulation system based on CT images].

OBJECTIVE: To design a new afterloading brachytherapy simulation system based on CT images. METHODS: This paper mainly focuses on the anthropomorphic pelvic phantom spiled by three pipelines and the nasopharyngeal carcinoma spiled by two pipelines. Microsoft Visual C++ was used to parse CT images for some information, then to reconstruct pipelines in the body of phantom or the patient and to give the three-dimensional coordinate of dwelling points. The dose distribution displayed on CT images was processed by the dose distribution calculation methods near single afterloading source and the dose optimization methods. VTK technology was used in the 3D display in the system. RESULTS: According to the reference points applied by doctors, the system can calculate reversely the dwelling time of dwelling points in pipelines and get satisfying dose distribution on CT images. Besides, it can reflect the 3D relationship between the dose volume and the normal tissues. CONCLUSIONS: This system overcomes some deficiencies of 2D afterloading brachytherapy simulation system based on X-ray films which are used widely in China. It supplies 3D display of dose distribution for clinical doctors. At present, the system is being tested in clinics.

lncRNA PCAT6 promotes non-small cell lung cancer cell proliferation, migration and invasion through regulating miR-330-5p.

BACKGROUND: Investigating the roles of lncRNA prostate cancer-associated transcript 6 (PCAT6) in modulating the growth and aggressiveness of non-small-cell lung carcinoma (NSCLC) cell. METHOD: The levels of PCAT6 in NSCLC tissues and cell lines were determined by quantitative real-time PCR assay. MTT as well as colony formation assays were applied to explore the effect of PCAT6 on the growth of NSCLC cell in vitro. Wound healing and Transwell assays were utilized to analyze the impact of PCAT6 on the migration and invasion of NSCLC cell. Bioinformatics analysis and luciferase reporter assay were used to prove that miR-330-5p was the target of PCAT6. Colony formation, wound healing, and Transwell invasion assays were applied to demonstrate that PCAT6 promoted NSCLC cell growth, migration, and invasion through binding miR-330-5p. Finally, xenograft model was used to explore the role of PCAT6 in the tumor growth of NSCLC cell in vivo. RESULTS: PCAT6 was highly overexpressed in NSCLC tissues and cells compared with normal tissues and non-tumorigenic bronchial epithelial cell line, BEAS-2B. Downregulation of PCAT6 markedly reduced the proliferation, migration, and invasion of NSCLC cell. Moreover, down-expression of PCAT6 significantly increased the level of miR-330-5p in NSCLC cell. Further functional experiments indicated that down-expression of miR-330-5p reversed the inhibitory effect of PCAT6 on NSCLC cell growth, migration, and invasion. CONCLUSION: Our results reveal that lncRNA PCAT6 facilitates the proliferation, migration, and invasion of NSCLC cell via competitively binding to miR-330-5p.

[Study on integrated Chinese and Western therapy and criterion for efficacy evaluation of gastroesophageal reflux disease--a clinical observation on 116 cases].

OBJECTIVE: To explore the criterion for efficacy evaluation of gastroesophageal reflux diseases (GERD) and the effect of treatment with integrated Chinese and Western medicine. METHODS: One hundred and sixteen patients of GERD were randomly assigned to two groups, group A treated with Western medicine and group B with integrated Chinese and Western medicine. Changes of symptoms, TCM syndromes, and endoscopic picture were observed before and after treatment. And the condition of recurrence was also inspected. RESULTS: The total effective rate was 84.5% and 67.2% in group A and B respectively. The long-term efficacy was better in group B than that in group A (P < 0.05), while there was no significant difference in short-term efficacy between the two groups. But the recurrence happened in both groups after withdrawal of treatment. The recurrence rate was significantly higher in patients of deficiency syndrome type and coexistence of deficiency and excess syndrome type than that in those of excess syndrome type (P < 0.05), and it was also higher in those whose pathogenesis was associated with mental factor (P < 0.05). In the other 20 patients the treatment was sustained for 10.4 +/- 11.4 months on average. There were 69.0% of the patients with normal esophagus mucosa shown by endoscope examination, and the pathological changes were not coincident with the symptoms and prognosis. CONCLUSIONS: Basically, no case of GERD could be cured, so, it is supposed that the item of "cure" in the criteria of therapeutic efficacy evaluation is advisably made over to "clinical control" or "obvious efficacy", and add an item of "no change" in the criterion. The importance of endoscopic picture for efficacy evaluation is doubtful and needs to be further discussed.

Magnetic Resonance Imaging for the Assessment of Long Bone Tumors.

BACKGROUND: Wide resection margins of osseous tumors are associated with a low incidence of local recurrence, making accurate measurement of the intraosseous extent of primary malignant long bone tumors is crucial. We compared the intraosseous tumor extent assessed by magnetic resonance imaging (MRI) with the gross specimen to evaluate the accuracy of MRI. METHODS: A total of 255 patients with primary malignant tumors in the long bones were included. Using MRI, we defined the length of tumor as the distance from the articular surface to the boundary between abnormal and normal marrow signal. The extent of the abnormal intraosseous signal was measured on unenhanced T1-weighted (T1WI) magnetic resonance images after chemotherapy. All gross surgical specimens were sectioned, and tumor extent was measured. Wilcoxon signed-rank test was used to test the differences between MRI and gross specimen findings. Spearman's correlation analysis was used to test the correlation between groups. RESULTS: Median tumor length by gross specimen (112 mm; range, 45-300 mm) was longer than that by MRI (108 mm; range, 45-304 mm; Z = -6.916, P < 0.001). Of 255 images, tumor length was accurately represented on 27 T1WI magnetic resonance images, overestimated on 79 images, and underestimated on 149 images. The median difference between imaging and gross specimen measurements was 2.0 mm (range: 1.0-15.0 mm) for the 79 cases where tumor length was overestimated, and 5.0 mm (range: 1.0-18.0 mm) for the 149 cases where tumor length was underestimated. The Spearman correlation demonstrated a high correlation of tumor length on gross specimen with the tumor length on MRI (R = 0.99, P < 0.01). CONCLUSIONS: We conclude that preoperative MRI could be a useful method in determining intramedullary malignant bone tumor boundaries and may serve as an accepted assessment method of long bone tumors before limb-sparing surgery.

Is There a Reliable Method to Predict the Limb Length Discrepancy after Chemotherapy and Limb Salvage Surgery in Children with Osteosarcoma?

BACKGROUND: For a child with osteosarcoma, prediction of the limb length discrepancy at maturity is important when planning for limb salvage surgery. The purpose of this study was to provide a reliable prediction method. METHODS: A retrospective review of Chinese children receiving chemotherapy for osteosarcoma before skeletal maturity was conducted. Standing full-length radiographs of the lower extremity were used for length measurements. Length-for-age curves were constructed using the LMS method. The lower limb multiplier for a specific age and gender was calculated using the formula M = Lm/L, where M was the gender- and age-specific multiplier, Lmwas the bone length at maturity, and L was the age-specific bone length. Prematurity and postmaturity radiographs were used to assess the accuracy of the prediction methods. RESULTS: A total of 513 radiographs of 131 boys and 314 radiographs of 86 girls were used to calculate the coefficients of the multiplier. The multipliers of 8-, 9-, 10-, 11-, 12-, 13-, 14-, 15-, 16-, 17-, and 18-year-old boys after chemotherapy for osteosarcoma were 1.394, 1.306, 1.231, 1.170, 1.119, 1.071, 1.032, 1.010, 1.004, 1.001, and 1.000, respectively; while for girls at the same ages, the multipliers were 1.311, 1.221, 1.146, 1.092, 1.049, 1.021, 1.006, 1.001, 1.000, 1.000, and 1.000, respectively. Prematurity and postmaturity femoral and tibial lengths of 21 patients were used to assess the prediction accuracy. The mean prediction error was 0 cm, 0.8 cm, and 1.6 cm for the multiplier method using our coefficients, Paley's coefficients, and Anderson's method, respectively. CONCLUSIONS: Our coefficients for the multiplier method are reliable in predicting lower limb length growth of Chinese children with osteosarcoma.