In order to study the inherent difference among terpenes producing yeasts from the point of metabolomics, we selected taxadiene producing yeasts as the model system. The changes of cellular metabolites during fermentation log phase of artificial functional yeasts were determined using metabolomics methods. The results represented that compared to W303-1A as a blank control, the metabolites in glycolysis, tricarboxylic acid cycle (TCA) cycle and several amino acids were influenced. And due to the changes of metabolites, the growth of cells was inhibited to a certain extent. Among the metabolites identified, citric acid content in taxadiene producing yeasts changed the most, the decreasing amplitude reached 90% or more. Therefore, citric acid can be a marker metabolite for the future study of artificial functional yeasts. The metabolomics analysis of taxadiene producing yeasts can provide more information in further studies on optimization of terpenes production in heterologous chassis.
Alkenes ; metabolism ; Amino Acids ; metabolism ; Citric Acid ; analysis ; Citric Acid Cycle ; Diterpenes ; metabolism ; Fermentation ; Glycolysis ; Metabolome ; Metabolomics ; Yeasts ; metabolism

BACKGROUND:COX-2 gene actual y exists in the joint fibroblast-like synoviocytes, it affects osteoarthritis occurrence and development. Understanding the differences of COX-2 gene expression levels at different stages of osteoarthritis synoviocytes has important theoretical significance for the occurrence and development of osteoarthritis, as wel as the role of synoviocytes in this process.
OBJECTIVE:To analyze the difference of COX-2 mRNA at different stages of osteoarthritis fibroblast-like synoviocytes.
METHODS:Synovial membrane from 44 osteoarthritis patients and 12 normal cases were selected. Primary cells were cultured to passage 4 fibroblast-like synoviocytes for the use in the experiment. COX-2 mRNA expression in osteoarthritis fibroblast-like synoviocytes and normal fibroblast-like synoviocytes was detected using real-time fluorescence quantitative RT-PCR. The relative quantitative analysis was performed using 2-ΔΔCt method.
RESULTS AND CONCLUSION:Expression of COX-2 mRNA in osteoarthritis fibroblast-like synoviocytes was significantly higher than that in normal fibroblast-like synoviocytes (P<0.05). The expression levels reached a peak at early osteoarthritis group, with significant differences compared with middle and late osteoarthritis groups (P<0.05). There was no significant difference between middle and later osteoarthritis groups (P>0.05). COX-2 mRNA might be important biological marker for the inflammation in osteoarthritis, and mainly plays a role in early osteoarthritis stage.

The key challenge to generate engineered cells by synthetic biology for producing 7-dehydrocholesterol (7-DHC) in a high titer is the match between functional module and chassis. Our study focused on solving this problem by combining different promoters and yeast chassis to increase 7-DHC production. To optimize the chassis in order to accumulate zymosterol, the substrate for 7-DHC synthesis, we overexpressed truncated HMG-CoA reductase (tHmglp) and squalene epoxidase (Erglp), both are key genes of yeast endogenous zymosterol biosynthetic pathway. In addition, we knocked out C-24 methyl transferase (Erg6p) and C-22 dehydrogenase (Erg5p) to inhibit the conversion of zymosterol to ergosterol. By introducing heterologous C-24 reductase under three promoters with different strengths, namely TDH3p, PGK1p and TDH1p, we constructed functional modules of diverse activities. Nine engineeredcells were generated based on the combination of these three modules and three chassis. The result shows that the engineered cell composed of functional module regulated by TDH3p and chassis SyBE_000956 had the highest 7-DHC production, indicating a better match than others. This study provides evidences for importance of match and empirical support for rational design of subsequent researches.
Cholesterol ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; Dehydrocholesterols ; metabolism ; Gene Knockout Techniques ; Hydroxymethylglutaryl CoA Reductases ; metabolism ; Industrial Microbiology ; Methyltransferases ; genetics ; Promoter Regions, Genetic ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; Synthetic Biology

Objective:To explore the effect of applying standardized patient teaching based on Calgary-Cambridge communication model in nurse patient communication training for junior college interns, and to enrich the teaching methods of nurse patient communication training in domestic medical institutions.Methods:This study was a quasi-experimental study. In July 2022, 78 students were selected from 335 junior college interns in the First Affiliated Hospital of Shandong First Medical University by random coding method, and were randomly divided into the experimental group (39 students) and the control group (39 students) by lot. The control group received routine training. The experimental group received standardized patient teaching based on Calgary Cambridge communication model: teaching the key points of communication, guiding demonstration based on Calgary Cambridge communication model, and guiding reflection and exploration. The nurse patient communication ability, nurse patient communication practice skills, communication self-efficacy, and teaching satisfaction of the two groups of interns were compared between the two groups after 8 weeks of training.Results:After training, the total score of nurse patient communication ability evaluation in the experimental group was (91.41 ± 5.35) points, higher than that in the control group (88.08 ± 7.40) points, there was significant difference ( t=2.24, P<0.05); after training, the communication self-efficacy score of the experimental group was (30.21 ± 4.28) points, higher than that of the control group (27.94 ± 5.09) points, there was significant difference ( t=2.09, P<0.05); the total score of communication practice skills in the experimental group was (173.59 ± 18.48) points, higher than that in the control group (158.44 ± 15.57) points, there was significant difference ( t=3.82, P<0.05); the total score of communication teaching and training satisfaction in the experimental group was (16.77 ± 2.94) points, higher than that in the control group (15.22 ± 1.90) points, and there was significant difference ( t=2.68, P<0.05). Conclusions:The standardized patient teaching based on Calgary Cambridge communication model can effectively improve the practical skills of nurse patient communication of junior college interns, and promote the improvement of their nurse patient communication self-efficacy, which is conducive to the improvement of nurse patient communication ability junior college intern.

Objective: Adiponectin (APN) is an endogenous cytokine that mediates the development and progression of various tumors through its receptors (AdipoRs). The present study aimed to detect the expression and distribution of APN and its receptors (AdipoR1 and AdipoR2) in tongue squamous cell carcinoma (TSCC). Moreover, we explored whether the locoregional expression of APN was reg-ulated by HIF-1α in the hypoxic microenvironment. Methods: The expression and distribution of APN and its receptors in TSCC tissues were analyzed by immunohistochemical. Lentiviral expression vector for HIF-1α shRNA was constructed and stably transfected in TSCC cells to knock down HIF-1α expression. The mRNA and protein expression levels of APN and its receptors were detected using RT-PCR and Western blot, respectively, after hypoxic treatment. Results: The locoregional expression of APN and AdipoR1, but not AdipoR2, was upregulated at the early stages of T1, T2, and/or N0 stage, respectively, in tumor tissues compared to that in control paracancer-ous tissues (P<0.05 or P<0.01). The expression of APN and AdipoR1, but not AdipoR2, in TSCC cells was up-regulated on hypoxic treat-ment. Moreover, the expression of APN and AdipoR1 was down-regulated after shRNA knockdown of HIF-1α under hypoxia. Conclu-sions: The APN-AdipoR1 signaling pathway was activated and regulated by HIF-1α in the hypoxic environment of TSCC tissues.

Osteochondral lesion of talus (OLT) is a foot and ankle disease characterized by ankle pain, which may impact the joint function and life quality. If managed improperly, it may lead to a further ankle arthritis, severely compromising the prognosis. The therapeutic effect of conservative treatment for OLT is still uncertain. Surgery is still the main treatment modality for OLT with various techniques. However, the optimized surgical technique is still inconclusive, furthermore, regeneration and repair of cartilage after debridement is also a great challenge for the treatment of OLT. Platelet-rich plasma (PRP) with good repair effect on cartilage injury is gradually applied in the treatment of OLT. However, there still lacks the unified understanding of the technique and specification of PRP for the treatment of OLT. Therefore, National Orthopedics Center of Shanghai Sixth People′s Hospital allied Foot Ankle Basic Research & Orthopedics Group, Chinese Association of Orthopedic Surgeons; Foot and Ankle Committee of Chinese Association of Sports Medicine Physicians; and Foot and Ankle Group of Orthopedic Specialized Branch of Shanghai Medical Association to organize related experts to formulate the Expert consensus on platelet- rich plasma treatment for osteochondral lesion of talus ( version2023). Fifteen recommendations were put forward upon PRP preparation, indications, contraindications and treatment methods of PRP for OLT, so as to standardize the PRP treatment for OLT.

Objective:To analyze the characteristics of COVID-19 case spectrum and spread intensity in different provinces in China except Hubei province.Methods:The daily incidence data and case information of COVID-19 were collected from the official websites of provincial and municipal health commissions. The morbidity rate, severity rate, case-fatality rate, and spread ratio of COVID-19 were calculated.Results:As of 20 March, 2020, a total of 12 941 cases of COVID-19 had been conformed, including 116 deaths, and the average morbidity rate, severity rate and case-fatality rate were 0.97/100 000, 13.5 % and 0.90 %, respectively. The morbidity rates in Zhejiang (2.12/100 000), Jiangxi (2.01/100 000) and Beijing (1.93/100 000) ranked top three. The characteristics of COVID-19 case spectrum varied from province to province. The first three provinces (autonomous region, municipality) with high severity rates were Tianjin (45.6 %), Xinjiang (35.5 %) and Heilongjiang (29.5 %). The case-fatality rate was highest in Xinjiang (3.95 %), followed by Hainan (3.57 %) and Heilongjiang (2.70 %). The average spread ratio was 0.98 and the spread intensity varied from province to province. Tibet had the lowest spread ratio (0), followed by Qinghai (0.20) and Guangdong (0.23). Conclusion:The intervention measures were effective in preventing the spread of COVID-19 and improved treatment effect in China. However, there were significant differences among different regions in severity, case-fatality rate and spread ratio.

Ketogulonigenium vulgare is an acid-producing strain in the process of two-step vitamin C fermentation. L-sorbosone dehydrogenase (SNDH) is one of the key enzymes during the biosynthesis of 2-keto-L-gulonic acid (2-KGA), the precursor of vitamin C. However, the catalytic mechanism of SNDH is unclear. According to the whole genome sequencing of K. vulgare, two genes encoding sorbosone dehydrogenases, one derived from the chromosome (named as sndhg) and one from plasmid (named as sndhp), were introduced into an industrial strain K. vulgare. The overexpression of gene sndhg had hardly effect on 2-KGA production, and the overexpression of gene sndhp produced an obvious byproduct in the fermentation broth. Combinational expression of sndhg/sndhp with pqqA (obtaining sndhg-pqqA and sndhp-pqqA modules) in K. vulgare resulted in the similar fermentation phenotype to two previous strains. After serial sub-cultivation of co-cultured Bacillus endophyticus with each engineered K. vulgare for 50 d, the conversion rate of 2-KGA increased by 15.4%, 179%, 0.65% and 125% compared with that of the parental K. vulgare with B. endophyticus. This study shows that adaptive evolution of microbial consortium is an effective strategy to increase the fitness between functional modules and chassis, thus quickly getting better strains for production of 2-KGA.
Aldehyde Oxidoreductases ; genetics ; metabolism ; Ascorbic Acid ; Bacillus ; Bacterial Proteins ; genetics ; metabolism ; Coculture Techniques ; Fermentation ; Industrial Microbiology ; Microorganisms, Genetically-Modified ; Rhodobacteraceae ; enzymology ; genetics ; Sugar Acids ; metabolism