Three-dimensional cell culture is widely recognized as a model in vitro that can better simu-late the interaction between tumor cells in vivo and cell microenvironment. Compared with the traditional two-dimensional cell culture,it has obvious advantages in the study of the observation of tumor cells biological be-havior,tumor resistance,radiation insensitivity,gene expression in vivo and in other aspects of the study. It can provide a valuable model in vitro for experimental study on the basis of tumor.

In recent years, Chinese medicine preparations have been applied in clinic increasingly with the development of tradi-tional Chinese medicine ( TCM) . Meanwhile, TCM-induced diseases and adverse reactions have attracted more and more attention. In the study, the main symptoms and etiological factors of TCM-induced diseases were summarized, and relative control methods and strat-egies were suggested. The summarization can provide theoretical support for clinical rational drug use, which shows clinical significance and scientific value for preventing TCM-induced diseases and reducing the frequency and severity of the diseases.

Objective:To establish a method which Nifedipine was used as an alternative control for determinating of three flavonoid glycosides in the tablets of Ginkgo biloba leaves by substitution method of reference substance.Methods:Make Nifedipine instead reference substance, the analysis was performed on a JADE-PAK C18 column (4.6 mm × 250 mm, 5 μm), with the mobile phase consisted of 0.5% phosphoric acid solution-methanol (adjusting pH value to 2.2) (50:50) at the flow rate of 1.0 ml/min, the column temperature was 35 ℃. The Nifedipine was selected as the reference substance of three flavonoid glycosides and the correction factor of Nifedipine. The content of total flavonoid glycosides was determined by relative correction factor f and substitute reference.Results:The linear ranges of Nifedipine, quercetin, kaempferide and isorhamnetin were 0.17-1.73 μg, 0.11-1.08 μg, 0.08-0.84 μg, 0.04-0.40 μg. The relative correction factor ( f) between Nifedipine and quercetin, kaempferide, isorhamnetin were 0.782 2, 0.768 5, 0.808 8, respectively. The average recovery ( n=6) were 98.48%, 98.03%, 94.55%. Conclusion:The results indicated that the HPLC method by substitute reference substance is economical, feasible and practical.

Objective:To further optimize the nutritional support program for elderly neurocritical patients, analyze the changes and influencing factors of serum albumin levels in elderly neurocritical patients after surgery, and explore the impact on prognosis.Methods:A retrospective cohort study was conducted to investigate the medical records of neurosurgical intensive patients in Shanghai Tenth People's Hospital from May 2015 to September 2019. All the patients were treated for at least 5 days after operation. Patients with severe liver and kidney dysfunction, mental illness and incomplete medical records were not included in the study. Patients aged ≥65 years were included as the experimental group and patients aged <65 years served as the control group. Generalized estimating equation and other models were used to analyze the difference in the trend of albumin level and its influencing factors between the two groups. Automated machine learning was used to explore the influence of factors such as albumin level, nutrition mode, age, sex, diagnosis, preoperative consciousness level on the prognosis of patients.Results:A total of 284 patients were included in the study, including 85 in the experimental group and 199 in the control group. There were no significant differences in baseline data such as sex, age, and proportion of nutritional patterns between the two groups. Generalized estimating equation and generalized mixed additive equation showed that albumin level in both groups reached the minimum on the 3rd day after operation, and the average level in the experimental group was significantly lower than that in the control group ( P=0.03). Cerebral hemorrhage, emergency surgery, and low preoperative albumin were risk factors ( P<0.05); intravenous supplementation of human albumin might have an adverse effect on the experimental group ( P = 0.047). Machine learning models suggested that the higher the serum albumin level at discharge, the better the prognosis at 1 month after surgery. The area under ROC curve was 0.906, the specificity was 93.7%, and the sensitivity was 67.0%. Conclusions:The changes of albumin level in elderly patients with neurological critical illness after surgery are basically the same as those in young patients, with the lowest albumin level on the 3rd day after surgery, and the albumin levels even lower on the 3rd day after surgery in patients with cerebral hemorrhage, emergency surgery and preoperative low albumin. The higher the albumin level at discharge, the better the prognosis of patients may be.

Objective To investigate the enhancing effect of Celecoxib on the anti-proliferative effect of cisplatin (DDP) against human cervical carcinoma HeLa cells and to explore the possible mechanism. Methods After treatment of HeLa cells with Celecoxib or/and DDP, the growth suppression of HeLa cells was measured by MTT assay. Cell cycle and apoptosis were examined by flow cylometry. Results The inhibition of Celecoxib on HeLa cells was dose -dependent and time -dependent.The inhibitory effect was remarkable enhanced in asynergistic or additive pattern after treatment with the combination of Celecoxib and DDP (P =0.000) . Celecoxib (50 μmol/L) and DDP (16 μmol/L) could respectively induce the apoptosis of HeLa cells and this effect of apoptosis induction was increased when combinated Celecoxib with DDP. They could change cell cycle of HeLa cells lines with G_0/G_1 phase cells increasing and S phase cells decreasing. Conclusion Celecoxib combined with DDP shows synergistic anti-tumor effects.

Based on the background of knowledge economy,factors which have affected the social status of a person can be divided into five levels: physical fitness,technical ability,mental capacity,potential energy and recombination energy.This article analyzes the educational style of medical students by using five-level-ability model which judges a person from the economic angle.It aims at providing constructive opinions on innovation,enhancement and improvement of modern medical education.

Objective To study the effects of hydrogen dioxide (oxygen free radical donator) and vitamin C (oxygen free radical scavenger) on the electric current of large conductance calcium -activated potassium channels (BKCa channels) in isolated outer hair cells in aging guinea pigs .Methods Acute enzyme was used to isolat outer hair cells of aging guinea pigs ,in which of BKCa channel's electric current was observed and recorded by whole-cell recording mode of patch -clamp .After recording the stable and normal electric current of BKCa channels ,added H2 O2 dilution (0 .2 mmol/L) 40 μl in the 2 ml chambers within freshly isolated outer hair cells so that the concen‐tration of H2 O2 in the balneum would be 4 μmol /L .The groups(n=5) received individually vitamin C solution (5 mg/ml) 0 ,10 ,20 ,40μl in the 2 ml chambers within freshly isolated outer hair cells so that the concentration of vi‐tamin C in the balneum would be 0 ,25 ,50 ,100 μg /ml ,observing and recording the effects of different concentration of vitamin C to electric current of BKCa channels .Results ①In the w hole-cell mode of patch -clamp ,the rapid activation and non-deactivation electric current with a string of large amplitude was recorded ,above -40~ -30 mV activation voltage .The electric current increased with the increasing membrane potential .The amplitude in‐creased continuously and performed characteristics of outward rectification .When the concentration of IbTX was 100 nmol /L ,the activity of the channel was completely blocked and confirmed BKCa channel's electric current .②Medication within three minutes ,when VT was +50 mV ,the BKCa channels'the maximum peak current densities of 4 μmol /L H2 O2 group rose from 22 .09 ± 0 .27 PA /PF to 43 .53 ± 1 .09 PA/PF ,amplification was 97 .06% .In H2 O2 4 μmol /L + vitamin C with different concentrations as 25 ,50 ,100 μg/ml groups ,the BKCa channels'elec‐tric current performed about concentration-dependent inhibition ,and electric current's amplitude and peak current density decreased with the increasing concentration of vitamin C ,the I-V curves were reduced .However ,this still could not be recovered to the normal levels .Conclusion The oxygen free radical /BKCa exists in the process .The vitamin C as oxygen free radical scavenger can reverse the process to a large extent .

Objective To investigate the toxicity of levodopa and dopamine on PC12 cells and neuroprotection of several anti-Parkinson drugs i.e. amantadine,pergolide and selegiline. Methods The possible cytotoxicity of levodopa and dopamine at different dosage on rat pheochromocytoma PC12 cells and the effects of some anti-Parkinson drugs (amantadine,pergolide and selegiline) on levodopa- or dopamine-induced cytotoxicity were determined by MTT assay and flow cytometry. Results There was a concentration- and time-dependent decrease in cell viability and a concentration-dependent increase in apoptotic cells induced by levodopa and dopamine ( P

To investigate the effect of losartan on the axis of prolylcarboxypeptidase (PRCP)--kallikrein of the two-kidney, one-clipped (2K1C) hypertensives rats, and explore the novel protection mechanism of losartan on the kidney. Sprague-Dawley (SD) rats were used to develop the 2K1C hypertensive rats. Then, the rats were treated with prazosin (5 mg x kg(-1) x d(-1)) or losartan (5, 15 and 45 mg x kg(-1) x d(-1)) or vehicle, separately. At the same time, the blood pressures were observed. After treated for four weeks, the ratio of right kidney weight and body weight, the change of glomerular morphology, and K+, Na+, creatinine and blood urea nitrogen (BUN) of the serum were used for evaluation of kidney. The expressions of PRCP mRNA in the kidneys were determined by RT-PCR. The protein levels of PRCP, tissue kallikrein, plasma kallikrein, TGF-beta1 in kidney or plasma were measured by Western blotting. Results showed that the changes of body weight and kidney weight ratio, glomerular fibrosis degree and the biochemistrical index of serum induced by hypertension were relieved when the hypertensive rats treated with losartan for four weeks. Meanwhile, treatment of losartan also significantly decreased expression of TGF-beta1 and increased expressions of PRCP, plasma kallikrein and tissue kallikrein. The protective effects of losartan on the kidney of 2K1C hypertensive rats are activation of the axis of PRCP-kallikrein and reducing the expression of TGF-beta1.

Objective:To establish a protein-protein interaction (PPI) network of 5 microRNA (miRNA) target genes differentially expressed in the plasma of coal-burning fluoride exposed population, and to screen genes and gene modules with important roles.Methods:Five miRNA (hsa-miRNA-3131, hsa-miRNA-4516, hsa-miRNA-6501-5p, hsa-miRNA-10b-5p, hsa-miRNA-4683) target genes differentially expressed in the plasma of coal-burning fluoride exposed population screened by our previous study were mapped to the STRING online database (https://string-db.org), and the PPI network was screened. The Cytoscape v3.6.0 software was used to visualize the PPI network, the topological attribute values degree and betweenness centrality were obtained by the NetworkAnalyzer plug-in, and the central node was filtered in the network (the node with the highest degree and the highest betweenness centrality). At the same time, the maximal clique centrality (MCC) analysis method in the CytoHubba plug-in was used to determine the important nodes in the PPI network. The cluster analysis was conducted by the MCODE plug-in, and the gene modules were screened in the PPI network. The protein names contained in the gene modules were submitted online to the KOBAS v3.0 database (http://kobas.cbi.pku.edu.cn/), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed on the gene modules selected by the MCODE plug-in.Results:The PPI network of target genes was consisted of 1 035 nodes and 4 346 edges. The degree (101) and betweenness centrality (0.010 723 89) of ubiquitin A-52 residue ribosomal protein fusion product 1 (UBA52) were the highest, which was the central node of the PPI network. According to MCC analysis, UBA52 was an important node in the PPI network. The top 5 gene modules were selected from the PPI network, and the highly enriched gene pathways in the KEGG pathway enrichment analysis of the 5 gene modules included ubiquitin mediated proteolysis, spliceosome, endocytosis, neuroactive ligand-receptor interaction and vesicular transport.Conclusion:The PPI network of 5 miRNA target genes differentially expressed in the plasma of people exposed to coal-burning pollution of fluoride is successfully established, and the UBA52 gene and the 5 main pathways of gene modules are selected.