In vitro cell experiment based on serum pharmacology is a common way to study the pharmacodynamic mechanism of Chinese medicine， and screening the optimal intervention concentration of serum containing Chinese medicine is a key step in the whole experiment. Over-diluted serum containing Chinese medicine or over-high concentration leads to false negative or positive results， while the optimal concentration of medicated serum reduces the number of groups in subsequent experiments as well as the operation error. Thus it is of great significance to screen the optimal serum concentration in studying the pharmacodynamic mechanism of Chinese medicine in in vitro cell experiments. However， there has been no unified standards for the screening methods at present. After reviewing the literature in China and abroad in the past 20 years， this paper conducted an analysis from three aspects of intragastric dose of Chinese medicine， blood collection time and serum dilution degree， and then summarized the screening methods for optimal concentration of serum containing drugs， providing guidance for the study of pharmacodynamic mechanism of Chinese medicine. The screening methods mainly included "same intragastric dose， same blood collection time， and different concentrations" "different intragastric doses， same blood collection time， and same concentration" "same intragastric dose， different blood collection time， and same concentration" "different intragastric doses， different blood collection time， and different concentrations" "serum lyophilized powder" "simulation of gradient concentration of serum containing western medicine" and "pure serum intervention". Among them， the former two were the dominant ways in current related research. The above screening methods had their own advantages and disadvantages， and researchers should make a reasonable choice according to the specific requirements and actual situation of the experiments.

Objective: To investigate the level of hypoxia inducible factor-1α (HIF-1α) on osteoblasts and angiogenesis-associated cytokines in bone marrow mesenchymal stem cells (BMSCs) from SD rats. Methods: BMSCs were isolated and cultured and identified by flow cytometry. Plasmid vectors containing upregulated and downregulated HIF-1α gene and a control vector were constructed. The plasmids were transfected into BMSCs by Lipofectamine®LTX transfection reagent, and the cells were divided into an overexpression experimental group, an overexpression control group, a low expression experimental group and a low expression control group. All components were stained with a lizarin red 3 d and 7 d after osteogenesis induction. The mRNA expression levels of the target gene HIF-1α, osteogenic differentiation-specific markers, including Runt-related transcription factor 2 (Runx2) and angiogenic markers, including platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-β (TGF-β), were detected by RT-PCR. Western blot was used to detect the protein expression of the target proteins HIF-1α, Runx2, and PDGF-BB. Results: The CD29- and CD45-positivity rates of BMSC surface markers identified by flow cytometry were 98.2% and 4.2%, respectively. RT-PCR results showed that the mRNA expression of HIF-1α, Runx2, TGF-β and PDGF-BB was observably increased (P < 0.001). The mRNA expression levels of HIF-1α, Runx2, TGF-β and PDGF-BB in BMSCs from the low expression experimental group were significantly reduced (P < 0.001). Western blot results showed that the expression levels of HIF-1α, Runx2 and PDGF-BB in BMSCs from the overexpression experimental group were all increased (P < 0.001). The expression levels of HIF-1α, Runx2 and PDGF-BB in BMSCs from the low expression experimental group were reduced (P < 0.001). Alizarin red staining results showed that the area of calcium nodules in the low expression experimental group was smaller than that in low expression control group, the area of red calcium nodules in the over expression experimental group was larger than that in over expression control group, and with the increase of osteogenic induction time, the calcification area of each group also increased. Conclusion Upregulation and downregulation of HIF-1α can regulate the osteogenic differentiation and the expression of angiogenesis related factors of BMSCs.

Purpose: We aimed to investigate whether obtaining a higher level of education was causally associated with lower breast cancer risk and to identify the causal mechanism linking them. Methods: The main data analysis used publicly available summary-level data from 2 large genome-wide association study consortia. Mendelian randomization (MR) analysis used 65 genetic variants derived from the Social Science Genetic Association Consortium as instrumental variables for years of schooling. The outcomes from the Breast Cancer Association Consortium (BCAC) were the overall breast cancer risk (122,977 cases/105,974 controls in women) and the two subtypes: estrogen receptor (ER)-positive breast cancer and ER-negative breast cancer. Fixed and random effects inverse variance weighted methods were used to estimate the causal effects, along with other additional MR methods for sensitivity analyses. Results: Results showed that each additional standard deviation of 4.2 years of education was causally associated with a 27% lower risk of ER-negative breast cancer (odds ratio, 0.73; 95% confidence interval, 0.64–0.84; p-value < 0.001). This finding was consistent with the results of the sensitivity analyses. Physical activities can help improve the protective effect of education against breast cancer, with relatively large mediation proportions. Education increases the risk of ER-positive breast cancer due to alterations in high-density lipoprotein level, triglyceride level, height, waist-to-hip ratio, body mass index, and smoking status, with relative medium mediation proportions. Other mediators including low-density lipoprotein, hip circumference, number of cigarettes smoked per day, time spent performing light physical activity, and performing vigorous physical activity for > 10 minutes explain a small part of the causal effect of education on the risk of developing breast cancer, and their mediation proportion is approximately 1%. Conclusion A low level of education is a causal risk factor in the development of breast cancer as it is associated with poor lipid profile, obesity, smoking, and types of physical activity.

Objective To explore the relationship between exposure to air pollutants and chronic kidney disease. Methods We searched and screened the literature on air pollutant exposure and CKD, using Pubmed, Medline, Embase, and Cochrane databases from inception to May 1, 2020. Chronic exposure to air pollutants and risk of chronic kidney disease were estimated. Results Air pollutants can cause kidney damage to varying degrees, and PM_2.5 and PM₁₀ can increase the risk of chronic kidney disease. CO, NO₂(NO_X) and SO₂ may increase the risk of chronic kidney disease. Conclusions Exposure to air pollutants, especially particulate matter( PM_2.5 and PM₁₀) ，is associated with an increased risk of chronic kidney disease.

Three dimensionally printed composite porous bone tissue engineering scaffolds have become a research focus. Composite polyvinyl alcohol (PVA) has good biocompatibilityand degradability, but it cannot be prepared indepen⁃dently because it cannot resist highmechanical resistance. This material shows many advantages, such as good biocom⁃patibility, degradability and mechanical properties, when compounded with other materials with good mechanical proper⁃ties and good biocompatibility. Therefore, 3D printed composite PVA scaffold material can optimize the performance of PVA scaffolds. This article reviews 3D printing bone scaffold technology, polyvinyl alcohol (PVA), and composite PVA scaffolds for in vivo and in vitro bone formation.

Zinc levels are high in pancreatic β-cells, and zinc is involved in the synthesis, processing and secretion of insulin in these cells. However, precisely how cellular zinc homeostasis is regulated in pancreatic β-cells is poorly understood. By screening the expression of 14 Slc39a metal importer family member genes, we found that the zinc transporter Slc39a5 is significantly down-regulated in pancreatic β-cells in diabetic db/db mice, obese ob/ob mice and high-fat diet-fed mice. Moreover, β-cell-specific Slc39a5 knockout mice have impaired insulin secretion. In addition, Slc39a5-deficient pancreatic islets have reduced glucose tolerance accompanied by reduced expression of Pgc-1α and its downstream target gene Glut2. The down-regulation of Glut2 in Slc39a5-deficient islets was rescued using agonists of Sirt1, Pgc-1α and Ppar-γ. At the mechanistic level, we found that Slc39a5-mediated zinc influx induces Glut2 expression via Sirt1-mediated Pgc-1α activation. These findings suggest that Slc39a5 may serve as a possible therapeutic target for diabetes-related conditions.

Objective: To analyze the quadrivalent influenza vaccine intention of 718 health care workers (HCWs) in the Pearl River Delta region from 2015 to 2017. Method: In May 2018, 718 HCWs from the department related to the diagnosis and treatment of influenza in 17 hospitals (6 tertiary hospitals, 5 secondary hospitals and 6 primary hospitals) from Guangzhou, Jiangmen, Zhuhai and Dongguan were selected by using stratified sampling method. Questionnaire survey and face-to-face interview were used to collect the information of influenza vaccination, the intention of the quadrivalent influenza vaccine, the acceptance of free and required vaccination policies, and recommendations for increasing influenza vaccination intentions from 2015 to 2017. The multivariate logistic regression was used to analyze factors associated with the vaccination intention. Results: A total of 718 HCWs were surveyed and 147 of them were interviewed face to face. Among them, the vaccination rate of primary hospitals [17.39%(40/230)] was higher than that of other hospitals (χ²=15.80, P<0.05). If the vaccine could be free, 84.82% (609/718) of HCWs would like to be vaccinated. The multivariate logistic regression showed that the factors, HCWs who were aged ≥50 years (OR=3.44, 95%CI:1.43-8.28), worked in department of prevention and health care (OR=2.35, 95%CI:1.16-4.75), learned about the quadrivalent influenza vaccine (OR=2.94, 95%CI:2.08-4.18), knowed that HCWs are priority (OR=2.33, 95%CI:1.56-3.48), and had a history of trivalent influenza vaccination from 2015 to 2017 (OR=4.70, 95%CI:3.08-7.15), were associated with the vaccination intention. Conclusion HCWs in the Pearl River Delta region had weak inclination of getting quadrivalent influenza vaccine. HCWs who were age (≥50 years old), worked in department of prevention and health care, learned about the quadrivalent influenza vaccine, knowed that HCWs are priority, and had a history of trivalent influenza vaccination from 2015 to 2017 were factors positively associated with the vaccination intention.

The development of orally administrated heparin drugs requires a systematic understanding of the interaction between heparin and gut flora. The in vivo distribution of fluorescein-labeled heparin that is orally administrated by mice was observed using fluorescein microscopy. In addition, the stability of heparin in simulated gastric and intestinal fluids, as well as the in vitro degradation of heparin by gut flora were detected by HPLC. The results show that orally administrated heparin was mainly distributed in the gastrointestinal tract of mice, and exerted structural stability under the condition of simulated gastric and intestinal fluids in vitro. However, heparin could be degraded by intestinal flora cultured in medium containing heparin. In order to further study the effect of orally administrated heparin on intestinal flora in mice, the fecal microbiota 16S rRNA fragment of C57BL/6J mice was tested by the Illumina Mi-Seq high-throughput sequencing technology. Compared with the gut flora of mice that orally administrated by saline, the biodiversity of gut flora in mice with orally administrated heparin was decreased. The difference of microflora structure was not significant at the phylum level, and the relative abundance of Alistipes, Parasutterella and Akkermansia was increased at the genus level, and the relative abundance of Bilophila, Enterorhabdus, Ruminiclostridium, Prevotellaceae_UCG_001, Ruminiclostridium-9, Bacteroides, Lachnoclostridium, Candidatus, Saccharimonas, Intestinimonas and Dubosiella was reduced. These findings indicate that heparin could influence the gut flora of mice. In addition, no obvious toxic and side effects were found in mice that orally administrated heparin, suggesting the safety of orally administrated heparin.
Animals ; Gastrointestinal Microbiome ; Heparin ; Mice ; Mice, Inbred C57BL ; RNA, Ribosomal, 16S

Objective: To investigate the expression of Formin-2（FMN2）protein in gastric cancer tissues and its correlation to clinicopathological features of gastric cancer patients, as well to explore its effect on the proliferation of gastric adenocarcinoma AGS cells. Methods: 84 cases of gastric adenocarcinoma tissues and corresponding para-cancerous tissues were surgically collected from patients treated in the First Affiliated Hospital of Air Force Military Medical University from September 2015 to September 2017. The expression of FMN2 in gastric adenocarcinoma tissues was detected by immunohistochemical staining and analyzed with RNA-Seq data-sets GEPIA. The relationship between FMN2 protein expression in gastric adenocarcinoma tissues and its clinicopathological features was also explored. MTT assay was used to detect the effect of FMN2 onAGS cell proliferation activity, and Western blotting was used to detect the effect of FMN2 on the expression of apoptosis-related protein caspase-3 in AGS cells. Results: The expression level of FMN2 in gastric adenocarcinoma tissues was significantly lower than that in matched adjacent tissues and the expression level of FMN2 was closely related to the TNM stage and differentiation of gastric adenocarcinoma (all P<0.05). Compared toAGS control group, the proliferation activity of AGS/FMN2 was significantly decreased and the expression of apoptosis-related gene Caspase-3 was markedly increased (all P<0.05). Conclusion: The expression of FMN2 was significantly decreased in gastric adenocarcinoma tissues and its low expression is closely related to the degree of tumor differentiation and clinical TNM stage. Moreover, FMN2 over-expression significantly decreased the proliferation of AGS cells. FMN2 may function as independent risk factor for the prognosis of gastric adenocarcinoma, which may provide new ideas for the treatment of gastric adenocarcinoma.

Objective: To investigate the expression of histone methyltransferase G9a in gastric cancer tissues and its correlation to prognosis, and to observe the effect of G9a inhibitor on the proliferation and apoptosis of gastric cancer cells. Methods: The expression level of G9a in gastric cancer tissues and its correlation to prognosis were analyzed by using the Kaplan-Meier Plotter and Oncomine database. Human gastric cancer cell line SGC-7901 and MKN-45 were selected as study subject. The expression level of G9a protein was detected by Western blotting. The morphological change of gastric cancer cells after the treatment of G9a inhibitor BIX01294 was observed. CCK-8 proliferation experiment and plate colony formation assay were used to examine the proliferation ability and clone formation rate of gastric cancer cells. The changes of cell apoptosis were detected by Annexin-V staining. Results: G9a was highly expressed in gastric cancer tissues (P<0.01), and the high expression of G9a was positively correlated with poor prognosis of gastric cancer patients (P<0.01). After the treatment of BIX01294, the morphology of gastric cancer cells was changed, the volume of gastric cancer cells reduced, the intercellular connections disappeared, and even the apoptotic manifestations appeared, such as the shrinking,, becoming round and cast-off etc. BIX01294 could significantly inhibit the proliferation and colony formation but promote the apoptosis of gastric cancer cells (all P<0.05). Conclusion: Histone methyltransferase G9a was highly expressed in gastric cancer tissues, and its high expression level was positively correlated with poor prognosis. The proliferation of gastric cancer cells was obviously inhibited while the apoptosis was significantly promoted after inhibiting G9a expression.