Objective: To investigate the effects of miR-200a on the proliferation of lung cancer cells and to identify its direct target genes. Methods:Real-time PCR was performed to analyze the miR-200a expression in 15 paired clinical specimens of non-small cell lung cancer and adjacent noncancerous tissues, human lung cancer cell lines (A549, NCI-H520, and SK-MES-1), and one human normal lung bronchial epithelial cell line (16HBE). The effects of miR-200a on the proliferation of A549 lung cancer cells were detected through CCK-8 method. The candidate target genes of miR-200a were identified by bioinformatics screening and then verified by dual luciferase reporter gene assay, real-time PCR, and Western blot. The effects of YAP1 downregulation on the proliferation of A549 lung cancer cell line were also observed through CCK-8 method. Results:The miR-200a expression in non-small cell lung cancer tissues and lung cancer cell lines was significantly decreased (P<0.01). The upregulation of miR-200a expression could significantly inhibit the pro-liferation of A549 lung cancer cells (P<0.01). Dual luciferase reporter gene indicated that miR-200a could directly affect the 3′-untrans-lated region of the YAP1 gene to inhibit luciferase activity (P<0.01). Real-time PCR and Western blot revealed that the upregulation of miR-200a expression could significantly reduce the mRNA and protein expression levels of YAP1 in A549 lung cancer cells (P<0.01). CCK-8 method indicated that the downregulation of YAP1 could significantly prevent the proliferation of A549 lung cancer cells (P<0.01). Conclusion:MiR-200a inhibits the proliferation of lung cancer cells by targeting YAP1. Thus, miR-200a elicits tumor suppression effects.

Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.
Agrobacterium ; Blotting, Southern ; Digestion ; Genome ; Genomics ; Methods ; Microscopy, Fluorescence ; Oxidoreductases ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Wounds and Injuries

In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: picking up a suitable amount of transformant's tissue (1–10 μg) to 20 μl 0.25% Lywallzyme solution, and vortexing for 10 s followed by incubation at 34 °C for 15 min. Finally, 2 μl of the suspension was used as templates to perform PCR and single target bands were successfully amplified from respective transformants of Tremella fuciformis, Pleurotus ostreatus, and Pleurotus tuber-regium. This procedure could be widely employed for screening transformants in mushroom transformation experiments.

Since the COVID-19 outbreak, medical institutions in different parts of the country have experienced personal protective equipment(PPE) shortages at different levels. The situation is especially severe in the COVID-19 epicenter. Overseas donation is one of the effective ways to ease the shortage of PPE. Based on the practice of Union Hospital of Tongji Medical College of Huazhong University of Science and Technology, the authors discussed the difficulties and risks for public hospitals to receive overseas donations, summarized experience and proposed suggestions which can be used as reference for other public hospitals and related departments.

This paper described the practice of medical cost control in Hubei's provincial state-owned hospitals. Their measures taken include combinations of overall planning and classification, total volume control and structural adjustment, external governance and internal management, as well as systematical management and difficulties breakthroughs. This paper proposed to further consolidate the outcomes of the abovementioned efforts by improving the fiscal compensation mechanism, furthering medical insurance payment reform, and continuing to monitor and publicize the medical expenses.

Objective To investigate the risk factors of acute kidney injury (AKI) occurring in patients with critical neurological disease, and the related factors affecting their prognosis. Methods The clinical data of 207 patients with critical neurological disease admitted to the Department of Critical Care Medicine of Anhui Provincial Hospital Affiliated to Anhui Medical University (South District) from January 2016 to March 2017 were analyzed retrospectively, they were assigned into an AKI group (40 cases) and a non-AKI group (167 cases), and according to the prognosis, the patients with AKI were subdivided into a survival subgroup (14 cases) and a death subgroup (26 cases). Clinical data of Glasgow coma score (GCS), acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), blood glucose, white blood cell count (WBC), central venous pressure (CVP), blood sodium, cystatin C, urea nitrogen (BUN) etc. index levels and the proportions of patients using glycerin fructose and furosemide before occurrence of AKI were collected. The indexes with statistical significant differences found in the univariate analysis were analyzed by multivariate logistic regression analysis to screen out the risk factors influencing the occurrence of AKI and the factors related to the prognosis of the AKI patients; the receiver operating characteristic curve (ROC) was drawn to assess the predictive value of risk factors in patients with severe neurological disease to develop AKI. Results The incidence of AKI was 19.3% (40/207) in the patients with critical neurological disease. The hospital mortality in AKI group was significantly higher than that in the non-AKI group [65.0% (26/40) vs. 22.2% (37/167), P < 0.01]. Compared with non-AKI group, GCS (4.44±1.65 vs. 5.39±1.62), CVP [cmH2O (1 cmH2O = 0.098 kPa): 7.69±2.66 vs. 8.98±2.56] were obviously lower in AKI group at admission, APACHEⅡ(24.50±3.67 vs. 20.05±4.42), blood glucose (mmol/L: 12.33±6.53 vs. 9.33±3.26), serum sodium (mmol/L: 144.75±10.85 vs. 140.58±5.23), WBC (×109/L: 16.15±6.25 vs. 12.79±4.22), Cystatin C (mg/L: 1.27±0.74 vs. 0.74±0.26) and BUN (mmol/L: 7.81±3.33 vs. 5.53±3.20) and proportion of male [77.5% (31/40) vs. 59.9% (100/167)], patients with the comorbidity of hypotension [37.5% (15/40) vs. 19.8% (33/167)], use of glycerin fructose [17.5% (17/40) vs. 3.6% (6/167)], or use of furosemide [70.0% (28/40) vs. 13.8%(6/167)] were significantly increased in AKI group, there was a statistically significant difference between the above two groups (all P < 0.05). Multivariate logistic regression analysis showed that the hyperglycemia [odds ratio (OR) = 1.201, 95% confidence interval (95%CI) = 1.01-1.42, P < 0.05] and use of furosemide for treatment (OR = 24.493, 95%CI =4.92-120.36, P < 0.01) were the independent risk factors for occurrence of AKI in critical neurological patients. ROC curve analysis showed that blood sugar had certain predictive value of developing AKI in patients with critical neurological disease, the area under the ROC curve (AUC) of blood glucose was 0.733, when the optimal cut-off value of blood glucose was 9.05 mmol/L, the sensitivity was 77.5% and the specificity was 62.6%. Compared with the survival subgroup in the patients with AKI, the GCS at admission in death subgroup was significantly lower (3.77±0.87 vs. 5.50±2.03), but their levels of blood glucose (mmol/L: 16.51±9.10 vs. 10.09±2.89) and BUN (mmol/L: 10.26±3.07 vs. 6.48±2.70) were obviously higher than those in the survival subgroup (all P < 0.05). Conclusion AKI is one of the common complications in patients with critical neurological disease, hyperglycemia and the use of furosemide are the independent risk factors of occurrence of AKI in such patients; the blood glucose has moderate predictive value; and lower GCS, higher glucose and BUN levels in AKI patients may enhance their risk of death.

Taking Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology as an example,it discusses the basic ideas and innovative practices of project organization from"passive order taking"to"active planning",platform construction from"free growth"to"directional cultivation",team construction from"wearing hats"to"focusing on actual combat",and achievement transformation from"resource guidance"to"comprehensive policy".It puts forward some suggestions that hospitals should play the leading role of"national team"in organizing scientific research and innovation practice,pay attention to the docking of national strategy,the linkage of university resources.

The development characteristics of tertiary public hospitals in China presented differently under the macro environment in different periods. Based on the theory of supply and demand chain of medical services, the authors discussed the key factors affecting the high-quality development of tertiary public hospitals under the new situation from the four elements of production, distribution, circulation and consumption, and put forward countermeasures and suggestions as follows: optimizing the supply of medical talents and system construction in the production process; speeding up the layout of high-quality resources and the construction of an integrated medical service system in the distribution process; strengthening imformation construction and the scientific and technological innovation in the circulation process; expanding healthcare/prevention integration and refining supporting policies in the consumption process, so as to provide reference for accelerating the high-quality development of public hospitals in China.

Since the COVID-19 epidemic broke out in Wuhan, medical institutions in different parts the country have experienced personal protective equipment (PPE) shortages at different levels. The situation is especially severe in the coronavirus epicenter. Overseas donation is one of the effective methods to ease the shortage of protective equipment. Based on the practice of Union Hospital of Tongji Medical College of Huazhong University of Science and Technology, the article discussed the difficulties and risks for public hospitals to receive overseas donations, summarized experience and proposed suggestions which can be used as reference for other public hospitals and related departments.

OBJECTIVE: The barks, leaves, and branches of Cinnamomum cassia have been historically used as a traditional Chinese medicine, spice, and food preservative, in which phenylpropanoids are responsible compounds. However phenylpropanoid biosynthesis pathways are not clear in C. cassia. We elucidated the pathways by descriptive analyses of differentially expressed genes related to phenylpropanoid biosynthesis as well as to identify various phenylpropanoid metabolites. METHODS: Chemical analysis, metabolome sequencing, and transcriptome sequencing were performed to investigate the molecular mechanisms underlying the difference of active components content in the barks, branches and leaves of C. cassia. RESULTS: Metabolomic analysis revealed that small amounts of flavonoids, coumarine, and cinnamaldehyde accumulated in both leaves and branches. Transcriptome analysis showed that genes associated with phenylpropanoid and flavonoid biosynthesis were downregulated in the leaves and branches relative to the barks. The observed differences in essential oil content among the three tissues may be attributable to the differential expression of genes involved in the phenylpropanoid and flavonoid metabolic pathways. CONCLUSION This study identified the key genes in the phenylpropanoid pathway controling the flavonoid, coumarine, and cinnamaldehyde contents in the barks, branches and leaves by comparing the transcriptome and metabolome. These findings may be valuable in assessing phenylpropanoid and flavonoid metabolites and identifying specific candidate genes that are related to the synthesis of phenylpropanoids and flavonoids in C. cassia.