The monosaccharide composition and the structure of polysaccharide from Pichia fermentans were analyzed. The yeast cells were broken with ultrasonic disruption and the crude polysaccharide was precipitated by addition of ethanol. Then, the crude polysaccharide was separated by diethylaminoethyl(DEAE) cellulose column separation and an acquisition of uniform water-soluble polysaccharide yeast polysaccharides 5(YP5) was obtained. The results of gas chromatographic analysis indicated that the glycosyl was made of rhamnose, arabinose, xylose, mannose, glucose and galactose, whose molar ratio was 9.51∶ 1.33∶ 2.31∶ 0.94∶ 84.93∶0.96, respectively. The molecular weight of polysaccharides YP5 was 8.3×10~4 measured by Ubbelohde viscosity method. Combination of IR and ~1H NMR analysis showed that the YP5 owned characteristic infrared absorption peaks of polysaccharide and contained amino groups. The β-D-glucan was main glucosidic linkage of polysaccharides PY5.

Objective To investigate the effects of up?regulated miR?206/miR?1 on the proliferation of breast cancer stem cells and the effect mech?anism. Methods Breast cancer stem cells(BCSCs)were isolated from breast cancer cell line MCF?7 by fluorescence?activated cell sorting. Cells in the experiment were divided into the blank control group,the negative control group,the miR?206 group and the miR?1 group. The BCSCs were transfected by negative control mimic,hsa?miR?206mimic and hsa?miR?1mimic in all groups except the blank control group. MiR?206and miR?1 expression levels as well as the transcription factor EVI?1 gene were detected by real time PCR. The expression levels of the transcription factor EVI?1 protein were detected by Western blot. MTT method was used to detect the effects of miR?206 and miR?1 on the proliferation of BCSCs. Results The BCSCs(CD44+/CD24-/low cells)isolated from MCF?7 cell lines were successfully cultured in serum?free medium for subsequent studies. After transfection of hsa?miR?206mimic and hsa?miR?1mimic for 48 hours,miR?206and miR?1relative expression levels increased. EVI?1mRNA ex?pression levels significantly decreased. The results of Western blot and MTT showed that up?regulated expression levels of miR?206 and miR?1 could significantly reduce the expression of EVI?1 protein and inhibited the proliferation of BCSCs. The differences in levels of miR?206,miR?1 and EVI?1 protein were statistically significant(P<0.05). Conclusion Up?regulated miR?206 and miR?1 expression can inhibit the proliferation ability of BCSCs,which may be related to the down?regulation of EVI?1.

Objective:To study the difference of the development between mandibular condylar cartilage and femoral head cartilage in vitro.Methods:Mandibular condyles and femoral heads were sampled from 1 2 neonatal mice and cultured in vitro.The samples before culture and after 6-week culture were examined by gross observation,HE staining,Alizarin Red staining and PCNA immunohistochem-istry respectively.Results:After in vitro culture,abnormal changes were observed in condyle cartilage,but the surface area of condyle cartilage was not changed(P >0.05).HE staining showed partial cartilage layer structure disappearance and the Alizarin Red staining confirmed calcification in the cartilage matrix.However,calcification was not found in femoral head cartilage,and the surface area of femoral head cartilage increased(P <0.05).HE staining showed the hypertrophied layer was thicker after culture than before and the Alizarin Red staining showed there was no calcification in the femoral cartilage matrix.The immunohistochemistry displayed PCNA posi-tive expression in both cartilage after culture.Conclusion:In vitro,the mandibular condylar cartilage matrix can be spontaneously cal-cificated.

Objective To study the rehabilitation effect of Bellidifolin for injuried sciatic nerve,and to explore whether ciliary neurotro-phic factor ( CNTF) is involved in this mechanism. Methods The right sciatic nerver of 225 male wistar rats was cut and sewed under mi-croscopy. Rats were devided into 5 groups,as control group,Bellidifolin 25 mg group,50 mg group、75 mg group and Mecobalamin group. The control group were injected sodium chloride,other groups were injected different dose of Bellidifolin and Mecobalamin. 1,3 and 5 weeks later, the motor nerve conduction velocity( MNVC) and gastrocnemius muscle cross-sectional area were detected,CNTF positive area were analysed by immunohistochemical method. Results There were differences among bellidifolin groups,control group and mecobalamin group in Nerve conduction velocity. Within Bellidifolin groups,50 mg group compared with 25 mg and 75 mg groups,there were statistically differences( P=0. 025). Three weeks after operation,gastrocnemius muscle cross-sectional area of control group,mecobalaming grop and Bellidifolin 25 mg group,50 mg group,and 75 mg group were(455. 06 ± 29. 38),(679. 03 ± 81. 48),(465. 31 ± 71. 55),(670. 24 ± 91. 26) and (669. 28 ± 78. 54) respectively,compared with control group and Bellidifolin 25 mg group,others had a significant difference(P<0. 05). CNTF expres-sion showed billidifolin 50 mg group are higher than others(P<0. 05). Conclusion Bellidifolin can improve the rehabilitation of injured sciatic nerve. CNTF is involved in this mechnism.

Objective To investigate Danhong in improving the movement function of reperfused rat after stroke. Methods Rats to make middle cerebral artery occlusion model,after 24 hours of reperfusion were divided into control group,high dose and low dose of Danhong groups and Venorruton group randomly. Rats in Control group were injected saline. High and low dose group were injected DanHong according to their weight,high dose as 14. 4 mg/kg,low dose as 3. 6 mg/kg. Venorruton group were injected Venorruton as 0. 04 g/kg. The infarcted square of rat brain was measured,the rats were tested with walking wood bar and two fore limb grasp strength. Results Rat infarcted squre in 24 hours and 3 days,control groups is the biggest,compared with others,there are statistically difference(P<0. 05). Score in each group of walking wood bar in 3rd days are (1. 30 ± 0. 91),(3. 78 ± 1. 72),(4. 18 ± 2. 05),(4. 63 ± 2. 45). Compared between control and other druge groups,there are statistically differnec(P<0. 05). There are statistically differences between control and other groups in two fore limb grasp testing(P<0. 05). Conclusion Danhong has the same effect which can improve the movement function to stroke rat as Venorruton.

Objective To explore the effects and mechanisms of prostaglandin E2 (PGE2) receptor 1 antagonist (SC-19220) on proliferation,prostaglandin synthase and extracellular regulated protein kinases (ERK) signal pathway induced by transforming growth factor β1(TGF-β1) in glomerular mesangial cells.Methods Mouse glomerular mesangial cells (GMCs) were divided into 5 groups:control group,TGF-β1 (10 μg/L) group,TGF-β1 (10 μg/L) plus SC-19220 group (0.1,0.5,1.0 μmol/L).The proliferation of GMCs was measured by CCK-8.The PGE2 in supernatant was measured by ELISA.The expression of connective tissue growth factor (CTGF),laminin (LN),cyclooxygenase 2(COX2),membrane-bound prostaglandin E2 synthase 1 (mPGES1) protein and mRNA was examined by Westem blotting and real-time quantitative PCR,ERK1/2 or phospho-ERK1/2 was measured by Western blotting as well.Results TGF-β1 induced the proliferation of GMCs and increased the secretion of PGE2.Besides,TGF-β1 significantly up-regulated the expression of CTGF,LN,COX2 and mPGES1 mRNA and protein (P ＜ 0.05),and increased the expression of phospho-ERK1/2 protein (P ＜ 0.05).However,SC-19220 significantly attenuated the changes of above-mentioned parameters and their activities (P ＜ 0.05).All the effects of SC-19220 were in dose-dependent manner.Conclusions SC19220 may reduce TGF-β1-induced cell damage by suppressing the activity of ERK1/2,and feedback inhibition of COX2,mPGES1 and PGE2,thus decreases the expression of LN and CTGF.

Purpose: During the major shift in China's policies on coronavirus disease 2019 (COVID-19), many residents will be infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) over a short period, including a few patients with breast cancer undergoing neoadjuvant chemotherapy (NAC). Moreover, it is unknown whether this comorbidity affects the efficacy of NAC for breast cancer and the patient's psychological state and quality of life (QOL). This study aims to answer these questions. Methods: The clinical data of 2,793 patients with breast cancer who received NAC at The Affiliated Hospital of Qingdao University were retrospectively collected. The infected and non-infected groups were divided according to whether they were infected with COVID-19 during NAC. Propensity score matching was used to reduce patient selection bias. The effectiveness, psychological well-being, and QOL of the two groups were compared. Results: No discernible differences were observed in the pathological complete response rates (p = 0.307) and major histological responses rate (p = 0.398) between the infected and non-infected groups. Following the full course of NAC, the Functional Assessment of Cancer Treatment General (p < 0.001) and Functional Assessment of Cancer Therapy for Breast Cancer (p < 0.001) were lower in the infected group than the non-infected group, the Hospital Anxiety and Depression Scale (HADS) anxiety scale (p < 0.001) and HADS depression scale (p < 0.001) were considerably higher in the infected group than the non-infected group. Conclusion With timely treatment and effective medical management, SARS-CoV-2 does not appear to affect the efficacy of NAC; however, it can significantly affect the QOL of patients and increase their psychological distress. Therefore, in addition to a timely assessment of the efficacy of NAC, it is necessary to dynamically understand the patient's psychological state and QOL.

DNA barcoding is a rapidly developing frontier technology in the world and will be useful in promoting the quality control and standardization of traditional Chinese medicine. Until now, many studies concerning DNA barcoding have focused on leaf samples but rarely on Chinese herbal medicine. There are three issues involved in DNA barcoding for traditional Chinese medicinal materials: (1) the extraction methods for total DNA of the rhizomes of the medicinal materials; (2) intra-specific variation among samples from different places of origin; (3) accuracy and stability of this method. In this study, Gentianae Macrophyllae Radix was used to verify the stability and accuracy of DNA barcoding technology. Five regions (ITS2, psbA-trnH, matK, rbcL, and ITS) were tested for their ability to identify 86 samples of Gentianae Macrophyllae Radix and their adulterants. After improving the DNA extraction method, genomic DNA from all samples was successfully obtained. To evaluate each barcode's utility for species authentication, PCR amplification efficiency, genetic divergence, and species authentication were assessed. Among all tested regions only ITS2 locus showed 100% of PCR amplification and identification efficiencies. Based on the established method, we successfully identified two samples of Gentianae Macrophyllae Radix bought in pharmacy to the original species.

Microbial metabolic engineering and synthetic biology are important disciplines of microbial technology nowadays. Microbial cells are fast growing, easy to be cultivated in large scale, clear in genetic background and convenient in genetic modification. They play an important role in many domains. Microbial cell factory means an artificial microbial metabolic system that can be used in chemical production. The construction of a microbial cell factory needs transferring of multiple genes or a whole metabolic pathway, which may cause some problems such as metabolism imbalance and accumulation of mesostates. This review focuses on the regulation strategies of different levels involving simultaneous engagement of multiple genes. Future perspectives on the development of this domain were also discussed.
Bacteria ; genetics ; Fungi ; genetics ; Gene Expression Regulation ; Gene Regulatory Networks ; genetics ; Genetic Engineering ; Industrial Microbiology ; Metabolic Engineering ; methods ; Metabolic Networks and Pathways ; genetics

The sigma-1 receptor is a molecular chaperone protein highly enriched in the brain. Recent studies linked it to many diseases, such as drug addition, Alzheimer's disease, stroke, depression, and even cancer. Sigma-1 receptor is enriched in lipid rafts, which are membrane microdomains essential in signaling processes. One of those signaling processes is ADAM17- and ADAM10-dependent ectodomain shedding. By using an alkaline phosphatase tagged substrate reporter system, we have shown that ADAM10-dependent BTC shedding was very sensitive to both membrane lipid component change and sigma-1 receptor agonist DHEAS treatment while ADAM17-dependent HB-EGF shedding was not; and overexpression of sigma-1 receptor diminished ADAM17- and ADAM10-dependent shedding. Our results indicate that sigma-1 receptor plays an important role in modifying the function of transmembrane proteases.
ADAM Proteins ; metabolism ; ADAM10 Protein ; ADAM17 Protein ; Amyloid Precursor Protein Secretases ; metabolism ; Animals ; Betacellulin ; COS Cells ; Cercopithecus aethiops ; Gene Expression ; HEK293 Cells ; Heparin-binding EGF-like Growth Factor ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Membrane Microdomains ; metabolism ; Membrane Proteins ; metabolism ; Receptors, sigma ; agonists ; metabolism