ObjectiveTo explore the molecular mechanism through which modified Guizhi Fulingwan （MGZFLW） promotes apoptosis and inhibits prostate cancer through the miR-34a-5p/δ-like protein 1 （DLL1）/Notch signaling pathway based on the traditional Chinese medicine theory of Yang transforming Qi while Yin constituting form. MethodsThe optimal intervention conditions of MGZFLW-containing serum and docetaxel （DXT） were screened by the cell counting kit-8 （CCK-8） method. Human prostate cancer cells （PC-3） were allocated into blank， MGZFLW （10%）， and DXT （30 nmol·L^-1） groups. The effects of MGZFLW-containing serum on the migration and proliferation of PC-3 cells were evaluated by cell scratch and colony formation assays， and the apoptosis of PC-3 cells was detected by terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL） assay. The cells were transfected with miR-34a-5p mimics and miR-34a-5p inhibitor. The mRNA levels of miR-34a-5p， DLL1， Notch1 and its downstream effector factors Notch1 intracellular domain （NICD1）， Hes family basic helix-loop-helix transcription factor 1 （Hes1）， and hairy/enhancer-of-split related with YRPW motif 1 （Hey1） were determined by Real-time PCR. The protein levels of DLL1， Notch1， NICD1， Hes1， and Hey1 were determined by a WES fully automatic protein system. The effect of miR-34a-5p on the DLL1-mediated Notch pathway was evaluated based on the results. Further， after MGZFLW-containing serum intervention， the expression of DLL1 and Notch1 was detected by the immunofluorescence （IF） assay， and the mRNA and protein levels of miR-34a-5p/DLL1/Notch pathway-related molecules were quantified by Real-time PCR and the WES fully automatic protein system， respectively， to explore the regulatory effect of MGZFLW-containing serum on the miR-34a-5p/DLL1/Notch pathway. ResultsMGZFLW-containing serum inhibited the viability， migration， and proliferation and promote the apoptosis of PC-3 cells （P<0.01）. The overexpression of miR-34a-5p down-regulated the mRNA and protein levels of key molecules in the DLL1/Notch signaling pathway and promoted cell apoptosis （P<0.01）， while inhibition of miR-34a-5p had the opposite effects. MGZFLW-containing serum reversed the activation of the DLL1/Notch pathway and the inhibition of apoptosis caused by miR-34a-5p inhibition， up-regulated the expression of miR-34a-5p， and down-regulated the expression of related molecules in this pathway （P<0.01）. ConclusionMGZFLW can up-regulate the expression of miR-34a-5p to inhibit the DLL1/Notch signaling pathway and promote the apoptosis of prostate cancer cells， thereby inhibiting prostate cancer. This study provides experimental evidence for clarifying the molecular mechanism of the anti-prostate cancer effect of this formula.

AIM:The aim of our study is to investigate the metabolic profile differences during cervical lesion progression and evaluate their potential clinical value in assisting the diagnosis of cervical cancer(CC).METHODS:Ul-tra-high-performance liquid chromatography coupled with high-resolution mass spectrometry(UHPLC-HRMS)was em-ployed to conduct non-targeted metabolomic analysis of cervical swab samples from 43 CC patients,34 high-grade squa-mous intraepithelial lesion(HSIL)patients,and 43 healthy controls.Based on the distinct features among the three groups,principal component analysis(PCA)was used to identify the metabolic differences among CC,HSIL and healthy groups.MetaboAnalyst 5.0 was then employed to perform KEGG pathway enrichment analysis on the differential metabo-lites.Finally,random forest machine learning algorithm was used to construct classification prediction models for distin-guishing CC from healthy,HSIL from healthy,and CC from HSIL.The performance of these models was evaluated using receiver operating characteristic(ROC)curve analysis.RESULTS:A total of 1 543 metabolites were identified across the healthy,HSIL and CC groups after filtration,with 407 metabolites differing between the groups.The study found that metabolite PGE2 was present in all three groups,with its expression levels progressively increasing with the progression of cervical lesions.Differential metabolite enrichment analysis demonstrated that CC is associated with specific cancer-relat-ed metabolic pathways,including the tricarboxylic acid cycle,tyrosine metabolism,tryptophan metabolism,and the pen-tose phosphate pathways.Additionally,the study developed three prediction models based on metabolic products for diag-nosing HSIL and CC:the full model,the simplified model,and the PGE2 model.The results indicated that metabolites ex-hibited strong diagnostic efficiency.Both the full model and the simplified model effectively distinguished CC from HSIL,CC from healthy,and HSIL from healthy.The AUC values for the full model were 0.90,0.92 and 0.84,respectively,while those for the simplified model were 0.81,0.95 and 0.85,respectively.Furthermore,the PEG2 model achieved AUC values of 0.74 and 0.80 for distinguishing CC from healthy and HSIL from healthy,respectively.CONCLUSION:The metabolic profiles of cervical cancer exhibit significant differences during the progression of cervical cancer,and these metabolites hold potential clinical value as biomarkers for cervical lesions.

AIM:The aim of our study is to investigate the metabolic profile differences during cervical lesion progression and evaluate their potential clinical value in assisting the diagnosis of cervical cancer(CC).METHODS:Ul-tra-high-performance liquid chromatography coupled with high-resolution mass spectrometry(UHPLC-HRMS)was em-ployed to conduct non-targeted metabolomic analysis of cervical swab samples from 43 CC patients,34 high-grade squa-mous intraepithelial lesion(HSIL)patients,and 43 healthy controls.Based on the distinct features among the three groups,principal component analysis(PCA)was used to identify the metabolic differences among CC,HSIL and healthy groups.MetaboAnalyst 5.0 was then employed to perform KEGG pathway enrichment analysis on the differential metabo-lites.Finally,random forest machine learning algorithm was used to construct classification prediction models for distin-guishing CC from healthy,HSIL from healthy,and CC from HSIL.The performance of these models was evaluated using receiver operating characteristic(ROC)curve analysis.RESULTS:A total of 1 543 metabolites were identified across the healthy,HSIL and CC groups after filtration,with 407 metabolites differing between the groups.The study found that metabolite PGE2 was present in all three groups,with its expression levels progressively increasing with the progression of cervical lesions.Differential metabolite enrichment analysis demonstrated that CC is associated with specific cancer-relat-ed metabolic pathways,including the tricarboxylic acid cycle,tyrosine metabolism,tryptophan metabolism,and the pen-tose phosphate pathways.Additionally,the study developed three prediction models based on metabolic products for diag-nosing HSIL and CC:the full model,the simplified model,and the PGE2 model.The results indicated that metabolites ex-hibited strong diagnostic efficiency.Both the full model and the simplified model effectively distinguished CC from HSIL,CC from healthy,and HSIL from healthy.The AUC values for the full model were 0.90,0.92 and 0.84,respectively,while those for the simplified model were 0.81,0.95 and 0.85,respectively.Furthermore,the PEG2 model achieved AUC values of 0.74 and 0.80 for distinguishing CC from healthy and HSIL from healthy,respectively.CONCLUSION:The metabolic profiles of cervical cancer exhibit significant differences during the progression of cervical cancer,and these metabolites hold potential clinical value as biomarkers for cervical lesions.

Selenium (Se) is an essential trace element for organisms. Se deficiency will cause diseases such as Keshan disease and Kashin-Beck in human being, and huge loss to animal husbandry. Currently available Se supplements have such problems as low Se content, poor bioavailability, and poor safety. Chlorella pyrenoidosa can produce bioavailable and safe organic Se under suitable conditions, which is thus a promising Se supplement. Therefore, in this study, we tried to improve the Se tolerance and accumulation of C. pyrenoidosa by directional adaptation. To be specific, we gradually increased the concentration of Na₂SeO₃ in medium to domesticate C. pyrenoidosa and optimized the adapting time and concentration gradient of Na₂SeO₃ during the adaptation. The results showed that the adapted C. pyrenoidosa was more tolerant to Se and had stronger Se enrichment ability. In 5 L fermenter, the adapted strains could tolerate 40 mg/L Na₂SeO₃ and the synthesis rate of organic Se was 175.6% higher. Then, Se addition method in the 5 L fermenter was optimized. The result demonstrated that addition of Na₂SeO₃ at 40 mg/L during heterotrophic culture achieved the final dry weight of C. pyrenoidosa cells at 106.4 g/L, content of organic Se at 1 227 mg/kg, and synthesis rate of organic Se at 1.36 mg/(L·h). Compared with the reported highest cell density of 75 g/L and the highest organic Se content of 560 mg/kg, the corresponding figures in this study were 41.9% and 119.1% higher, respectively. In conclusion, directional adaptation can remarkably improve the Se tolerance and enrichment of C. pyrenoidosa.
Animals ; Humans ; Selenium/pharmacology* ; Chlorella ; Heterotrophic Processes

OBJECTIVE: To analyze the characteristics of cervical microecology in late reproductive-age women with cervical lesions and explore new methods for preventing cervical lesions. METHODS: Cervical smears were obtained from a total of 147 women of late reproductive age, including 24 with high-risk HPV infection (HR-HPV), 27 with low-grade squamous intra-epithelial lesions (LSIL), 36 with high-grade squamous intra-epithelial lesions (HSIL), 35 with cervical cancer (CC) and 25 healthy women. llumina MiSeq sequencing of V3-V4 region of the 16S rRNA gene amplicons was used to characterize the vaginal microbiota of the women. OTUs analysis of the valid data was performed, and the α-diversity (Chao1, Simpson's Index and Shannon Index) and β-diversity (T-test, weighted UniFrac β diversity, and MetaStat analysis) were evaluated. RESULTS: Dilution curve and species accumulation boxplot validated the quality of the samples. OTUs analysis of the 5 groups demonstrated that cervical bacterial genus consisted primarily of CONCLUSIONS The abundance of
Female ; Humans ; Microbiota ; Papillomaviridae ; Papillomavirus Infections ; RNA, Ribosomal, 16S/genetics* ; Uterine Cervical Neoplasms ; Vaginal Smears

The main functional ingredients of hot water extract of Chlorella pyrenoidosa (CPE) were investigated through a bioassay-guided fractionation based on free radical scavenging and macrophage proliferation effects. The main functional ingredients of CPE were polysaccharides (PS) that were isolated by high pressure extraction, Sevag method, ethanol precipitation and ultrafiltration separation. Crude polysaccharides were further separated and purified by ion exchange chromatography DEAE52 and size exclusion chromatography Sephadex G-100. The purified fractions were analyzed by gel permeation chromatography. Molecular weights of the purified fractions PS-1-4-2, PS-1-3-2 and PS-2-3-3 were 3.97×10⁴, 2.28×10⁴ and 4.1×10³ Da, respectively. Bioassay-guided fractionation results indicated that CPE could remove free radicals and promote Ana-1 cells proliferation, mainly due to its various components working together. The components of free radicals scavenging mainly concentrated in PS-1-3, PS-1-4, PS-2-3 and PS-2-4. The components of Ana-1 proliferation mainly concentrated in PS-1-3, PS-1-4 and PS-2-3. This study established the activity screening method of main functional component from CPE, and got three new functional ingredients. It can be used to guide the development of high value products, further promote the industrialization process of microalgae energy, and realize microalgae 'high value products, microalgae energy and microalgae carbon' integration of exemplary role.

Objective To compare the dose, clinical efficacy and acute adverse reactions of intensity modulated radiotherapy(IMRT)and three-dimensional conformal radiotherapy(3D-CRT)combined with three-dimensional brachytherapy (3D-BT) in the treatment of concurrent radiotherapy and chemotherapy for advanced stage cervical cancer patients. Methods Data collection was performed from January 2011 to November 2015 in Chinese PLA General Hospital and Inner Mongolia Cancer Hospital.All 89 patients with advanced stage (Ⅱb-Ⅲb) cervical cancer were treated by pelvic radiotherapy and concurrent chemotherapy, 46 cases of them received IMRT and 3D-BT(IMRT group), 43 cases received 3D-CRT and 3D-BT(3D-CRT group),along with cisplatin chemotherapy.The dose accumulation of external beam radiotherapy and 3D-BT was calculated by deformable image registration to analyze clinical efficacy, acute adverse reactions and prognosis of the two groups.Results (1)Dose of radiotherapy:planning target volume(PTV)coverage of IMRT group and 3D-CRT group were respectively(95.4±4.7)% and(95.1±5.1)%, without significant differences (t=0.289, P=0.773). Compared with the patients treated with 3D-CRT, the volumn receiving at least 30 Gy (V30), V50of rectum, colon, bladder and small intestine and V20of bone marrow in the IMRT group were significantly decreased (P<0.05). Regarding the combined dose, the maximum dose (Dmax) and the minimum dose received by the most exposed 2 cm3volume of the analyzed organ(D2CC)of rectum,colon,bladder and small intestine of IMRT group were significantly lower than those of 3D-CRT group (P<0.05). (2) Short-term efficacy: the effective rate of IMRT and 3D-CRT group were respectively 93% (43/46) and 91% (39/43), with no significant differences (χ2=0.237,P=0.626). (3) Acute adverse reactions: compared with 3D-CRT, IMRT could significantly reduce grade 1-2 acute toxicity in gastrointestinal [63%(29/46) vs 84%(36/43)], genitourinary [17%(8/46) vs 37%(16/43)] and hematologic [57%(26/46) vs 79%(34/43)] system (all P<0.05). There were no significant differences of grade 3 acute adverse reactions of gastrointestinal,genitourinary and hematologic system between two groups(all P>0.05). No grade 4 acute adverse reactions were observed. (4) Prognosis: the overall survival rate at 1, 2-year of IMRT and 3D-CRT group were respectively 95.6%,89.1% and 93.1%,86.1%.The progression-free survival rateat 1, 2-year of IMRT and 3D-CRT group were 91.1%, 89.1% and 88.4%, 86.1%, respectively. There were no significant differences in overall survival rate and progression-free survival rate between two groups (P>0.05). Conclusions Compared with 3D-CRT, IMRT combined with 3D-BT has dosimetry advantages based on dose accumulation algorithms by deformable image registration. IMRT could ensure clinical efficacy and significantly reduce the incidence rate of acute toxicities.

This research was aimed at establishing the pilot-scale purification technology of lipopeptide from marine-derived Bacillus marinus. We studied lipopeptide surfactivity interferences on scale-up unit technologies including acid precipitation, methanol extraction, solvent precipitation, salting out, extraction, silica gel column chromatography and HZ806 macroporous absorption resin column chromatography. Then, the unit technologies were combined in a certain order, to remove the impurities gradually, and to gain purified lipopeptide finally, with high recovery rate throughout the whole process. The novel pilot-scale purification technology could effectively isolate and purify lipopeptide with 87.51% to 100% purity in hectograms from 1 ton of Bacillus marinus B-9987 fermentation broth with more than 81.73% recovery rate. The first practical hectogram production of highly purified lipopeptide derived from Bacillus marinus was achieved. With this new purification method, using complex media became possible in fermentation process to reduce the fermentation cost and scale-up the purification for lipopeptide production. For practicability and economy, foaming problem resulting from massive water evaporation was avoided in this technology.

Chlorella is nutritious and has been used as a functional food much earlier than the other microalgae. C. pyrenoidosa, the potential microalgae which is currently cultured and developed for the new strategic industry of biofuels production and biological CO2 fixation, is a new resource food announced by the Ministry of Health of the People's Republic of China late 2012. Accumulation of high value-added substances in C. pyrenoidosa during the cultivation for lipid makes it possible to reduce the costs for C. pyrenoidosa-based biofuels production. Among these potential substances, hot water extract of Chlorella (CE), commercially known as "Chlorella growth factor", is the unique one that makes Chlorella more precious than the other algae, and the market price of CE is high. It is believed that CE is effective in growth promotion and immunoregulation. However, there is no systematic analysis on the research status of CE and its bioactivity. The present report summarized recent research progress of CE and its bioactivity. Generally, besides the main effect on immunoregulation and tumor inhibition, CE was efficient in improving metabolic syndrome, scavenging for free radicals, protecting against ultraviolet damage, chelating heavy metals, and protecting liver and bowel. Several major challenges in CE research as well as its prospects were also analysed in the present report.
Biofuels ; Chlorella ; chemistry ; Humans ; Lipids ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Water

Microalgae is a single-cell organism with the characteristics of high light energy utilization rate, fast growth rate, high-value bioactive components and high energy material content. Therefore, microalgae has broad application prospects in food, feed, bioenergy, carbon sequestration, wastewater treatment and other fields. In this article, the microalgae biotechnology development in recent years were fully consulted, through analysis from the literature and patent. The progress of microalgal biotechnology at home and abroad is compared and discussed. Furthermore, the project layout, important achievements and development bottlenecks of microalgae biotechnology in our country were also summarized. At last, future development directions of microalgae biotechnology were discussed.
Bibliometrics ; Biofuels ; Biomass ; Biotechnology ; trends ; Microalgae ; metabolism ; Waste Water