As one of the key enzymes in cell metabolism, the activity of citrate synthase 3 (CS3) regulates the substance and energy metabolism of organisms. The protein members of CS3 family were identified from the whole genome of apple, and bioinformatics analysis was performed and expression patterns were analyzed to provide a theoretical basis for studying the potential function of CS3 gene in apple. BLASTp was used to identify members of the apple CS3 family based on the GDR database, and the basic information of CS3 protein sequence, subcellular localization, domain composition, phylogenetic relationship and chromosome localization were analyzed by Pfam, SMART, MEGA5.0, clustalx.exe, ExPASy Proteomics Server, MEGAX, SOPMA, MEME, WoLF PSORT and other software. The tissue expression and inducible expression characteristics of 6 CS3 genes in apple were determined by acid content and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Apple CS3 gene family contains 6 members, and these CS3 proteins contain 473-608 amino acid residues, with isoelectric point distribution between 7.21 and 8.82. Subcellular localization results showed that CS3 protein was located in mitochondria and chloroplasts, respectively. Phylogenetic analysis divided them into 3 categories, and the number of genes in each subfamily was 2. Chromosome localization analysis showed that CS3 gene was distributed on different chromosomes of apple. The secondary structure of protein is mainly α-helix, followed by random curling, and the proportion of β-angle is the smallest. The 6 members were all expressed in different apple tissues. The overall expression trend from high to low was the highest relative expression content of MdCS3.4, followed by MdCS3.6, and the relative expression level of other members was in the order of MdCS3.3 > MdCS3.2 > MdCS3.1 > MdCS3.5. qRT-PCR results showed that MdCS3.1 and MdCS3.3 genes had the highest relative expression in the pulp of 'Chengji No. 1' with low acid content, and MdCS3.2 and MdCS3.3 genes in the pulp of 'Asda' with higher acid content had the highest relative expression. Therefore, in this study, the relative expression of CS3 gene in apple cultivars with different acid content in different apple varieties was detected, and its role in apple fruit acid synthesis was analyzed. The experimental results showed that the relative expression of CS3 gene in different apple varieties was different, which provided a reference for the subsequent study of the quality formation mechanism of apple.
Citric Acid ; Malus/genetics* ; Citrate (si)-Synthase ; Phylogeny ; Citrates

BACKGROUND:Several studies have found that the total flavonoids of rhizome drynariae can promote neovascularization in the induced membrane,improve the biological properties of the induced membrane,and accelerate bone remodeling in the induced membrane,but the related molecular mechanisms still need to be further explored. OBJECTIVE:To explore the effect of total flavonoids of rhizome drynariae on bone remodeling in rat femoral Masquelet-induced membrane model by regulating H-type blood vessels. METHODS:Thirty-six male Sprague-Dawley rats were stratified by body mass and then randomly divided into blank group,model group and traditional Chinese medicine group,with 12 rats in each group.A 4-mm femoral bone defect model was established in all the rats.Bone defects in the model group and traditional Chinese medicine group were filled with polymethylmethacrylate bone cement.At 6 weeks after modeling,the tail bone of the rats was implanted in the blank group,as well as in the other two groups after removal of bone cement.The traditional Chinese medicine group was given 157.5 mg/kg per day of total flavonoids of rhizome drynariae at 3 days after bone implantation,while the model and blank groups were given the same amount of saline by gavage until the 8th week after bone implantation.Bone graft samples were taken for relevant testing at 8 weeks after implantation. RESULTS AND CONCLUSION:X-ray films showed that in the blank group,the fracture line in the defect area was clear,and only a small amount of bone callus formed;in the model group,the bone defect area still existed,where discontinuous cortical bone was visible;in the traditional Chinese medicine group,the defect area was filled with newborn bone tissues,the bone marrow cavity and part of the cortical bone formed,and the fracture line disappeared.Micro-CT scans showed that the amount of new bone in the defect area was low in the blank group,the number of bone trabeculae in the defect area was significantly increased in the model group,and a large amount of new bone tissue was filled in the bone defect area in the traditional Chinese medicine group.Hematoxylin-eosin staining results showed that in the blank group,only a small amount of new bone formed in the defect area and the quality of osteogenesis was poor;in the model group,there was more new bone tissue in the defect area,but some fibrous connective tissues were interspersed within the bone tissue;and in the traditional Chinese medicine group,a large amount of new bone formed in the defect area and the quality of osteogenesis was the best.CD31/Emcn immunofluorescence double-labeling staining results showed that the number of H-type blood vessels in the newborn bone tissue in the bone defect area of the blank group was sparse and sparsely distributed;compared with the blank group,there were more H-type blood vessels in the bone tissue in the bone defect area of the model group,and the blood vessels were distributed in relatively regular strips;the number of H-type blood vessels in the bone defect area of the traditional Chinese medicine group was the highest and the blood vessels were densely distributed.To conclude,the total flavonoids of rhizoma drynariae can upregulate the expression of H-type blood vessels to enhance the angiogenic-osteogenic effect,improve the osteogenic efficiency of the rat femoral Masquelet induced membrane model,and promote bone remodeling.

OBJECTIVE: To develop a drug-loaded composite microsphere that can simultaneously release the berberine (BBR) and naringin (NG) to repair infectious bone defects. METHODS: The NG was loaded on mesoporous microspheres (MBG) to obtain the drug-loaded microspheres (NG-MBG). Then the dual drug-loaded compound microspheres (NG-MBG@PDA-BBR) were obtained by wrapping NG-MBG with polydopamine (PDA) and modifying the coated PDA with BBR. The composite microspheres were characterized by scanning electron microscopy, X-ray diffraction, specific surface area and pore volume analyzer, and Fourier transform infrared spectroscopy; the drug loading rate and release of NG and BBR were measured; the colony number was counted and the bacterial inhibition rate was calculated after co-culture with Staphylococcus aureus and Escherichia coli for 12 hours to observe the antibacterial effect; the biocompatibility was evaluated by live/dead cell fluorescence staining and cell counting kit 8 assay after co-culture with rat's BMSCs for 24 and 72 hours, respectively, and the osteogenic property was evaluated by alkaline phosphatase (ALP) staining and alizarin red staining after 7 and 14 days, respectively. RESULTS: NG-MBG@PDA-BBR and three control microspheres (MBG, MBG@PDA, and NG-MBG@PDA) were successfully constructed. Scanning electron microscopy showed that NG-MBG@PDA-BBR had a rough lamellar structure, while MBG had a smooth surface, and MBG@PDA and NG-MBG@PDA had a wrapped agglomeration structure. Specific surface area analysis showed that MBG had a mesoporous structure and had drug-loading potential. Low angle X-ray diffraction showed that NG was successfully loaded on MBG. The X-ray diffraction pattern contrast showed that all groups of microspheres were amorphous. Fourier transform infrared spectroscopy showed that NG and BBR peaks existed in NG-MBG@PDA-BBR. NG-MBG@PDA-BBR had good sustained drug release ability, and NG and BBR had early burst release and late sustained release. NG-MBG@PDA-BBR could inhibit the growth of Staphylococcus aureus and Escherichia coli, and the antibacterial ability was significantly higher than that of MBG, MBG@PDA, and NG-MBG@PDA ( P<0.05). But there was a significant difference in biocompatibility at 72 hours among microspheres ( P<0.05). ALP and alizarin red staining showed that the ALP positive area and the number of calcium nodules in NG-MBG@PDA-BBR were significantly higher than those of MBG and NG-MBG ( P<0.05), and there was no significant difference between NG-MBG@PDA and NG-MBG@PDA ( P>0.05). CONCLUSION NG-MBG@PDA-BBR have sustained release effects on NG and BBR, indicating that it has ideal dual performance of osteogenesis and antibacterial property.
Rats ; Animals ; Osteogenesis ; Delayed-Action Preparations/pharmacology* ; Microspheres ; Berberine/pharmacology* ; Anti-Bacterial Agents/pharmacology* ; Escherichia coli

Objective:To analyze the effect of problem-based learning (PBL) combined with case-based learning (CBL) and clinical pathway (CP) teaching methods in standardized residency training in department of hepatobiliary surgery.Methods:A total of 64 residents who received the standardized residency training in the Department of Hepatobiliary Surgery in Shaanxi Provincial People's Hospital from July 2018 to July 2019 were selected and divided into the observation group and the control group. The control group used PBL + CBL teaching methods, while the observation group adopted PBL + CBL + CP teaching methods. The after-department examination scores and the teaching cognition scores of the two groups were compared. SPSS 15.0 was used for t-test and Chi-square test. Results:The after-department examination scores of the two groups were compared. Compared with the control group, the examination scores of professional theories, case analysis and operation skills in the observation group were significantly higher, and the difference was statistically significant ( t = 6.98, 7.85, 7.01, P < 0.05). In terms of recognition of teaching, the observation group was significantly higher than the control group, and the difference was statistically significant ( t = 9.14, P < 0.05). Conclusion:The PBL + CBL + CP teaching is conducive to the comprehensive and systematic mastery of knowledge and the rapid establishment of scientific clinical thinking. It has a strong scientific and systematic nature and is worthy of promotion.

Objective: To systematically evaluate the clinical effect of Xiaoke Decoction in the treatment of type 2 diabetes. Methods: Chinese databases such as CNKI, Wanfang, Weipu Chinese Biomedical Journal Database, and Chinese Medical Biological Literature Database, PubMed, Cochrane Library, Embase, and Web of Science were searched for English language literature from their inception until November 2019. A Meta-analysis was performed using RevMan 5.3 and Stata 12.0. Results: Thirty-eight studies were included in this study, with a total of 3757 patients. It was found that adding Xiaoke Decoction could improve total efficiency. The Xiaoke Decoction groups surpassed the western medicine groups regarding improvement in total efficiency (OR = 3.49; 95% CI: 2.78–4.39, P < 0.00001). Adding Xiaoke Decoction could lower the fasting plasma glucose (FPG) level. The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in FPG levels (MD = −1.14; 95% CI: −1.36 to 0.92, P < 0.00001). Adding Xiaoke Decoction could lower the 2 h postprandial blood glucose (2hPG) level. The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in 2hPG (MD = −1.40; 95% CI: −1.61 to 1.19, P < 0.00001). Adding Xiaoke Decoction could lower glycated hemoglobin (HbA1c). The Xiaoke Decoction groups surpassed the western medicine groups regarding reduction in HbA1c (MD = −0.77; 95% CI: −0.95 to 0.58, P < 0.00001). It was found that adding Xiaoke Decoction could lower the traditional Chinese medicine (TCM) syndrome score. The TCM syndrome scores among patients in the Xiaoke Decoction group were lower than those among patients in the control group after treatment (MD = −4.90; 95% CI: −7.22 to 2.57, P < 0.0001). At the same time, we conducted a subgroup and sensitivity analysis of age and intervention duration on the heterogeneity of total efficiency, FPG, 2hPG, HbAlc, and TCM syndrome score outcome indicators. For detecting publication bias, an egger test was conducted. Conclusion: Compared with western medicine alone, Xiaoke Decoction has more advantages for the treatment of type 2 diabetes with respect to total efficiency, FPG, 2hPG, HbAlc, and TCM syndrome score.

Objective:To analyze the influence of lifestyle changes on body weight and metabolic parameters during the early stage of corona virus disease 2019 (COVID-19) epidemic and regular epidemic prevention period in physical examination population.Methods:A total of 801 subjects from Nanjing enterprises and institutions who underwent physical examination in the Department of Health Promotion Center, the First Affiliated Hospital of Nanjing Medical University in May of 2019 to 2021 were included in this study. The basic information and data of body mass index, fasting blood glucose, total cholesterol, low density lipoprotein cholesterol (LDL-C), triglyceride and uric acid were collected. Information about dietary and exercise habits was obtained through pre-examination questionnaires. The data of 2019 was set as baseline data, data of 2020 represented information during the early stage of COVID-19 epidemic and data of 2021 represented information in regular epidemic prevention period. The subjects were divided into underweight group, normal weight group, overweight group, and obese group according to the body mass index at baseline. The Friedman test was applied to compare changes in body mass index and metabolic parameters across the population. Chi-square test was used to compare changes in dietary and exercise habits. Multivariate logistic regression analysis was adopted to explore the influencing factors of body mass index changes.Results:During the early stage of COVID-19 epidemic, the proportion of weight gain was the highest in the underweight group (42.9%), the proportion of weight loss was the highest in the overweight group (24.2%), and the obese group has the most stable body weight (70.6%) ( P=0.004). Men ( OR=0.56, 95% CI: 0.36-0.87) were less likely to gain weight than women ( P=0.010). There was no significant differences in weight change among all groups in the regular epidemic prevention period ( P=0.380). During the early stage of COVID-19 epidemic, the levels of fasting blood glucose, total cholesterol and LDL-C were significantly lower than those of baseline [4.98 (4.66, 5.42) vs 5.23 (4.91, 5.66) mmol/L, 4.98 (4.36, 5.67) vs 5.11 (4.54, 5.77) mmol/L, 2.90 (2.45, 3.33) vs 3.23 (2.77, 3.74) mmol/L], and the uric acid level was higher [333.0 (275.5, 397.0) vs 311.0 (257.5, 368.0) μmol/L] (all P<0.001). In regular epidemic prevention period, the levels of body mass index, fasting blood glucose, total cholesterol and LDL-C were significantly higher than those in the early stage of the epidemic [24.0 (21.9, 26.3) vs 23.8 (21.7, 26.1) kg/m 2, 5.18 (4.85, 5.62) vs 4.98 (4.66, 5.42) mmol/L, 5.12 (4.42, 5.76) vs 4.98 (4.36, 5.67) mmol/L, 3.06 (2.59, 3.57) vs 2.90 (2.45, 3.33) mmol/L], while the uric acid was significantly lower [319.0 (265.0, 377.0) vs 333.0 (275.5, 397.0) μmol/L] (all P<0.001). During the early stage of the epidemic, the reduction proportion of unhealthy diet in the home group was significantly higher than that in the outing group (19.5% vs 11.4%), and the increment proportion of exercise in the outing group was significantly higher than that in the home group (5.1% vs 1.6%) (both P<0.05). In regular epidemic prevention period, the increase rate of unhealthy diet in the home group was significantly higher than that in the outing group (26.8% vs 13.0%) ( P<0.001), and there was no significant difference in exercise between the two groups ( P=0.325). During the early stage of COVID-19 epidemic and in the regular epidemic prevention period, unhealthy diet>3 times per week ( OR=3.85, 3.01, 95% CI: 1.74-8.51, 1.41-6.39) was positively correlated with weight gain, and regular exercise ( OR=4.35, 2.61, 95% CI: 2.05-9.23, 1.15-5.91) was positively correlated with weight loss (all P<0.05). Conclusions:During the early stage of COVID-19 epidemic and in the regular epidemic prevention period, the lifestyle in the physical examination population has an impact on body weight and metabolic indicators. In the early stage of the epidemic, unhealthy diet and exercise decreased, and metabolic indicators such as blood glucose and lipids decreased. People with low body weight tend to gain weight. In the regular epidemic prevention period, the subjects′ exercise increased but unhealthy diet also increased, and blood glucose, lipid and body weight elevated significantly.

Objective:To compare cerebral vascular hemodynamic indexes (CVHI) and revised Framingham stroke risk profile (FSP) in predicting the risk of stroke among physical examiners.Methods:A total of 1 049 individuals were enrolled from the Health Management Center, Jiangsu Province Hospital during January 2019 and January 2020. Correlation analysis was carried out to confirm the correlations of risk factors of stroke with CVHI accumulative score and FSP. Multivariate logistic regression analysis was performed to evaluate the influence of CVHI classification, hypertension, diabetes and smoking on the 10-year risk of stroke evaluated by FSP.Results:In this study, the number of people whose CVHI accumulative score was less than 75 was 163 (15.54%) and the number of people whose 10-year probabilities of stroke was greater than or equal to 6% was 202 (19.26%). The CVHI accumulative score was negatively related with FSP ( r=-0.284, P<0.001). The result demonstrated that the CVHI classification, hypertension, diabetes and smoking were independent predictors for the 10-year risk of stroke evaluated by FSP. Conclusions:The proportion of individuals with stroke risk in medical examination population evaluated by CVHI is lower than by FSP. Combination of CVHI and FSP can effectively predict the risk of stroke among physical examiners.

Objective: To explore the clinical therapeutic method and effect of the thumb-tack needle for subcutaneous embedding therapy on treating insomnia with insufficiency of heart and spleen among breast cancer patients. Methods: Totally 80 breast cancer patients with insomnia with insufficiency of heart and spleen treated in department of oncology, the Second Affiliated Hospital of Xi′an Jiao Tong University from March 2018 till November 2018 were selected and assigned into an experiment group and a control group by random number table, 40 cases in each one. The experiment group applied thumb-tack needle for subcutaneous embedding therapy, selecting Shenmen, Sanyinjiao, Zusanli, Anmian as major acupoint and Taibai as assistant acupoint. The control group used fake needles at the same places. After four-week treatment, we evaluated the therapeutic effect using the Pittsburgh Sleep Quality Index (PSQI) and observed side effects. Results: Before treatment, no significantly statistical differences were observed between the two groups on global and each dimension′s PSQI scores (P>0.05). After treatment, the PSQI scores and PSQI global score of the experiment group on sleep quality, sleep latency, sleep duration, sleep efficiency, sleep disturbances, and daytime dysfunction were (1.58±0.53), (1.72±0.33), (1.57±0.64), (1.61±0.68), (1.05±0.42), (1.27±0.53), (8.91±2.73) points respectively, which were (1.86±0.71), (1.90±0.41), (1.85±0.47), (1.97±0.83), (1.42±0.91), (1.56±0.45), (10.86±4.53) points respectively in the control group. There was significant difference between the two groups on PSQI global scores (t=1.998-2.637, P<0.05). The number of patients in the experiment group assessed as clinical cure, obviously effective, effective and noneffective were 8, 13, 9, 10 respectively, which were 1, 5, 12, 22 in the control group respectively. The clinical efficacy showed significant difference between the two groups (χ²=27.68, P<0.01). No side effects, such as infection, needle drop-off and etc. were observed. Conclusions The thumb-tack needle for subcutaneous embedding therapy could improve sleep quality of breast cancer patients with insomnia with insufficiency of heart and spleen, and it′s easy to operate, safe with little side effects.