This study aimed to investigate the ameliorative effect of Xinyang Tablets on myocardial fibrosis in uremic cardiomyopathy(UCM) using single-cell sequencing technology. UCM mouse models were established by 5/6 nephrectomy(NPM) and randomly divided into the model group, Xinyang Tablets group, and sham-operated(sham) group as the control. The Xinyang Tablets group received postoperative interventions of Xinyang Tablets(0.34 g·kg~(-1)). After eight weeks, the hearts of the mice in each group were disassociated and subjected to 10×Genomics single-cell sequencing. The data were subjected to t-SNE dimensionality reduction, K-means clustering, and CellMarker annotation prior to analyzing differential expression and cell differentiation trajectories using the Seurat and Monocle3 tools. Additionally, the CellChat tool was used to parse intercellular signaling communication. The results showed that a total of nine types of cells including fibroblasts, endothelial cells, and immune cells were identified in this study. The single-cell expression results of fibroblasts and Gene Ontology(GO) enrichment analysis showed that Xinyang Tablets regulated myocardial fibrosis factors and related signals. Mimetic timing analysis identified three major differentiation trajectories of mouse cardiac fibroblasts and identified the expression of secreted phosphoprotein 1(Spp1) as consistent with the fibroblast differentiation trajectory. Cellular interaction network analysis showed that the communication signals between mouse cardiac fibroblasts and other cells were weakened in the Xinyang Tablets group compared with the model group. The results of ligand-receptor interaction analysis showed that the interaction between myeloid cell-derived osteopontin(OPN) and cardiac fibroblasts and between myeloid cell Spp1 ligand and cardiac fibroblast receptor of mice in the Xinyang Tablets group was weakened compared with the model group. In conclusion, Xinyang Tablets may improve myocardial fibrosis in UCM by inhibiting both endogenous and exogenous OPN at the single-cell level.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Cardiomyopathies/pathology* ; Single-Cell Analysis ; Male ; Fibrosis/drug therapy* ; Myocardium/metabolism* ; Uremia/metabolism* ; Tablets ; Mice, Inbred C57BL ; Humans

OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Bronchopulmonary Dysplasia/epidemiology* ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Premature ; Infant, Newborn ; Pregnancy ; Respiratory Distress Syndrome, Newborn/epidemiology* ; Retrospective Studies ; Treatment Outcome

Objective: To investigate the independent and joint effects of chronotype and sleep duration on self-rated health in medical students. Methods: A cross-sectional study was conducted in 1 526 medical students selected through proportional stratified cluster random sampling from a medical university in Zhejiang province. A questionnaire survey was conducted to collect the information about their basic demographic characteristics, chronotype, sleep duration, and other lifestyle factors such as midnight snack, sedentary behavior, physical activity, meal time, and self-rated health. The independent and joint effects of chronotype and sleep duration on self-rated health were assessed by logistic regression model after controlling for confounding variables. Results: The numbers of the students with evening chronotype, neutral chronotype, and morning chronotype were 664 (43.5%), 442 (29.0%), and 420 (27.5%), respectively. Among the medical students, 42.8% (653) had poor self-rated health. Compared with those with the morning chronotype, the adjusted ORs for those with neutral chronotype and evening chronotype were 1.69 (95%CI: 1.23-2.31) and 2.43 (95%CI: 1.81-3.26), respectively, trend test P<0.001. Compared with those with sleep duration of 8 h or above per night, the adjusted ORs for those with sleep duration of 7 and ≤6 h per night were 1.40 (95%CI: 1.07-1.84) and 2.38 (95%CI: 1.69-3.37), respectively, trend test P<0.001. In the joint effect, compared with those with the morning chronotype and sleep duration of 8 h or above per night, the adjusted OR for those with evening chronotype and sleep duration of ≤6 h per night was 6.53 (95%CI: 3.53-12.09). Conclusions: Both evening chronotype and insufficient sleep were associated with increased odds of poor self-rated health in medical students, and they had joint effects. Therefore, it is necessary to promote early to bed, early to rise and adequate sleep in medical student to maintain their health.
Humans ; Students, Medical ; Cross-Sectional Studies ; Sedentary Behavior ; Sleep

Objective To investigate the effect of microRNA-9-5p (miR-9-5p) regulating transient receptor potential melastatin 7 (TRPM7) on myocardial ischemia-reperfusion (MIR) in rats. Methods Thirty-two SD rats were divided into sham operation group, model group, miR-9-5p overexpression group and empty vector control group. The MIR model was established by ligation of left coronary artery. The sham operation group was not ligated. miR-9-5p agomir and agomir NC were injected into tail vein 24 hours before model establishment in miR-9-5p overexpression group and empty vector control group. The myocardial injury was observed by HE staining. The expression of miR-9-5p was detected by Real-time PCR. The serum levels of interleukin(IL)-6, tumor necrosis factor alpha(TNF-α), IL-1β, creatine kinase isoenzyme MB (CK-MB), cardiac troponin Ⅰ (cTnI), lactate dehydrogenase (LDH) and the contents of malondialdehyde (MDA) and superoxide dismutase (SOD) in myocardium were measured were measured by ELISA. Cardiomyocyte apoptosis was detected by TUNEL. Double luciferase assay verified the relationship between miR-9-5p and TRPM7. The protein expressions of TRPM7, Bcl-2, Bcl-2 associated X (Bax), phosphorylated nuclear factor kappa-B 65 (p-NF-κB p65) and toll like receptor 4 (TLR4) were detected by Western blotting. Results The expression of miR-9-5p was low in myocardial tissue of rats (P<0.05). Overexpression of miR-9-5p could reduce the expression levels of CK-MB, cTnI and LDH, and improve the degree of myocardial injury. Compared with the model group, the apoptosis rate, Bax protein expression, MDA, IL-6, TNF-α and IL-1β contents in myocardial cells of miR-9-5p overexpression group decreased, while Bcl-2 protein expression and SOD content increased (P<0.05). The result of dual luciferase assay showed that TRPM7 was the target gene of miR-9-5p, and the protein expressions of TRPM7, p-NF-κB p65 and TLR4 in miR-9-5p overexpression group were lower than those in model group (P<0.05). Conclusion MiR-9-5p can inhibit myocardial cell apoptosis, oxidative stress and inflammation induced by myocardial ischemia-reperfusion, and inhibit TLR4/NF-κB pathway by regulating TRPM7.

OBJECTIVE: To explore the mechanism of acupuncture plus medication on treatment of Alzheimer's disease (AD). METHODS: Sixty adult SD rats were randomly divided into a normal group, a sham operation group, a model group, an electroacupuncture (EA) group, a gastrodin group and an EA+gastrodin group, 10 rats in each one. The rat model of AD was established by intraperitoneal injection of D-galactose and bilateral hippocampal injection of Aβ1-40. Two weeks after modeling, the rats in the EA group and EA+gastrodin group were treated with EA at "Baihui" (GV 20) "Dazhui" (GV 14) and bilateral "Zusanli" (ST 36), 30 min per treatment, once a day for consecutive 4 weeks. The rats in the gastrodin group and EA+gastrodin group were treated with intraperitoneal injection of gastrodin, once a day for consecutive 4 weeks. The rats in the normal group, model group and sham operation group were not treated. The morphology of hippocampal neurons was observed by using HE staining. The expression of Bcl-2 and Bax in the hippocampal CA1 area was detected by using immunohistochemical method. The expression of Bcl-2 and Bax protein in hippocampus was detected by using Western blot. RESULTS: The HE staining results showed the arrangement of neurons in the hippocampal CA1 area was regular in the normal group and the sham operation group, and the cytoplasm and nucleus were clearly visible. The neurons in the model group were severely damaged; the cell arrangement was not close, and the cell morphology was incomplete. Compared with the model group, the cell morphology of each intervention group was significantly improved. The immunohistochemistry results showed that, compared with the normal group and the sham operation group, the expression of Bcl-2 in the hippocampal CA1 region in the model group was decreased (<0.05), but the expression of Bax was enhanced (<0.05); compared with the model group, the expression of Bcl-2 was increased (all <0.05) and the expression of Bax was decreased (all <0.05) in all intervention group; compared with the EA group or the gastrodin group, the expression of Bcl-2 was enhanced (<0.05) and the expression of Bax was decreased (<0.05) in the EA+gastrodin group. The result of Western blot method was consistent with that of immunohistochemistry method. CONCLUSION EA and gastrodin could significantly inhibit the expression of Bax and up-regulate the expression of Bcl-2, and the combination of EA and gastrodin has the most significant effect. This indicates that EA combined with gastrodin has synergistic effect on inhibiting the apoptosis of neurons in hippocampus in AD rats, which may be one of the mechanisms of EA plus medication on AD lesions.
Alzheimer Disease ; Animals ; Electroacupuncture ; Hippocampus ; Proto-Oncogene Proteins c-bcl-2 ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein

OBJECTIVETo investigate the differences of metabolic pathways of leucocyte-deplated RBCs prepared by using lipid whole blood and nomal blood during routine storage so as to provide some reference for clinical blood use.

METHODSTwenty U whole blood from 20 donors, including 10 U lipid blood and 10 U normal whole blood, were selected for preparing leukodepleted red blood cells, red blood cells were taken from storage bags on day 0, 14 and 35, respectively. Metabolites in the red blood cells were analyzed, red blood cell metabolic extracts were detected by UPLC-MS/MS. The metabolite data of RBC from 2 groups were analyzed by SIMCA-P 13.0 software using OPLS-DA and by SPSS 19.0 using Mann-Whitney U test. Difference of metabolic pathways was described according to different metabolites.

RESULTSThe glucose, adenine, pyruvic acid, GSH, GSSG and niacinamide levels on day 0 in lipid RBCs were higher than those in the control group(P<0.05). The glucose, pyruvic acid and GSH levels on day 14 in lipid RBCs were lower than those in the control group (P<0.05), and the levels of adenine, GSSG and niacinamide were higher than that in the control group (P<0.05). The glucose level on day 0 was lower than that in the control group (P<0.05), and the levels of adenine and niacinamide were higher than those in the control group (P<0.05). but the pyruvic acid, GSH and GSSG levels were not significantly different between 2 groups (P>0.05).

CONCLUSIONCompared with the normal red blood cells, the energy metabolism pathway decreases in lipid red blood cells within the storage period and pentose phosphate pathway increases.

OBJECTIVETo investigate the effects of non-surgical periodontal therapy on serum inflammatory factors and metabolism levels in obese rats with experimental periodontitis.

METHODSSixteen obese rats with experimental periodontitis were randomly divided into treatment group and control group with non-surgical periodontal therapy and no treatment, respectively. Oral glucose tolerance test was performed before treatment and 2 weeks after the treatment. All the rats were sacrificed 2 weeks after treatment and the orbital vein blood was taken to detect fasting blood glucose, fasting insulin, and serum level of C-reactive protein (CRP). Results Two weeks after periodontal treatment, fasting blood glucose (t=2.445, P=0.034) and beta cell function index (t=-2.543, P=0.027) were significantly lower in the treatment group than in the control group. Compared with those in the control group, CRP level (t=2.388, P=0.028) and the area under the curve in the oral glucose tolerance test (t=12.053, P=0.000) decreased significantly in the treatment group.

CONCLUSIONNon-surgical periodontal treatment can reduce serum CRP level and improve glucose metabolism in obese rats.

OBJECTIVETo investigate the effect of periodontal therapy in controlling periodontitis and on insulin resistance and lipid metabolism in obese rats with periodontitis.

METHODSSprague-Dawley rats were randomized into normal group (group C), obese group (group O), periodontitis combined with obesity group (group P) and periodontal treatment group (group T). The obese rats in groups P and T were subjected to ligation of the maxillary second molar with silk thread to induce experimental periodontitis, and the rats in group T received periodontal therapy after the ligation. All the rats were sacrificed at the age of 24 weeks for measurement of blood lipids, insulin and blood glucose levels, and insulin resistance index (HOMA-IR) was calculated. The expressions of insulin receptor substrate-1 (IRS-1) and IRS-2 in the liver tissues were detected using real-time quantitative polymerase chain reaction (RT-PCR).

RESULTSCompared with the obese rats in group O, the rats in group P showed significantly higher HOMA-IR and LDL-C and lower expressions of IRS-1 and IRS-2 mRNA expression and HDL-C level (P<0.05). Compared with those in group P, the mRNA expressions of IRS-1 and IRS-2 and HDL-C level were significantly increased and LDL-C level, TC level and HOMA-IR were all decreased in group T (P<0.05), but the level of TG was comparable between the two groups. Pathological examination revealed lessened inflammatory cell infiltration and tissue destruction in the upper jaw of the rats in group T; the rats in group P presented with the most obvious upper jaw destruction and steatosis and inflammatory cell infiltration in the liver.

CONCLUSIONPeriodontal inflammation can downregulate the expression of IRS-1 and IRS-2 and increase insulin resistance and dyslipidemia in obese rats. Periodontal therapy produces a beneficial effect in improving insulin resistance and reducing dyslipidemia in obese rats.

BACKGROUND: As a blood concentrate rich in growth factors and fibrin, platelet-rich fibrin (PRF) is able to significantly promote bone regeneration. PRF mixed with bone substitutes is commonly used to repair bone defects in oral maxillofacial region. But it remains controversial whether PRF as a sole material in sinus floor elevation can produce enough new bone tissues.OBJECTIVE: To evaluate the osteogenic potential of PRF as a sole grafting material in sinus floor elevation by means of an animal model.METHODS: Twelve healthy adult mongrel dogs aged 12-18 months were selected and randomized into three groups (groups A, B and C), with four dogs in each group. Upper first molars on both side of each dog were extracted 12 weeks prior to sinus floor elevation and simultaneous implants placement. Different materials were used to fill in the space between the sinus membrane and implant. Groups A, B and C were filled with PRF, autologus bone particles and autologus blood clot respectively. After 12 weeks, specimens were harvested to analyze the new bone formation by gross observation, X-ray examination and histological evaluation.RESULTS AND CONCLUSION: New bone formation was found to embrace all the implants close to the bony sinus floor, and the tips of all implants were free from bone coverage. The new bone height of groups A, B and C were (3.135±0.288),(3.218±0.345), and (1.898±0.157) mm, respectively. The new bone density of groups A, B and C were (65.06±5.88),(75.34±8.18), and (56.92±4.95) g/cm3, respectively. There were significant differences between the new bone height in groups A and C as well as in groups B and C (P < 0.05). Also, significant differences were found between the new bone density in groups A and B, B and C, A and C (P < 0.05). Within the confine of this experiment, it is feasible to use PRF as a sole grafting material in the sinus floor elevation to generate new bone, but the new bone volume is limited.

Objective To analyze the chemical components of the ethanol-soluble portion of Liuwei Dihuang Decoction,and to explore its mechanism in kidney-Yin deficiency of mice.Methods Bioactive components were analyzed by HPLC,the method was used at Hypersile C1s column(4.6 mm × 250 mm,5 μm),the mobile phase consisted of methanol-water (27 ∶ 73),the detective wavelength was 236 nm,the flow rate was 1.0 mL · min-1,the column temperature was 25 ℃.The mice were divided randomly into three groups (n =10):blank group,model group,experimental group.The mice were gavaged with oral administration of corresponding medicines (9.75 g · kg-1)for nine weeks.At time 6 d after administration,in addition to that the normal group was given distilled water,the other groups were gavaged with hydrocortisone injection per day according to 50 mg · kg-1 body weight for 4 d to establish the kidney Yin deficiency mice model.The contents of cyclic adenosine monophosphate (cAMP),cyclic guanosine monophosphate (cGMP) were determined by ELISA and the follicle-stimulating hormone (FSH),estradiol (E2),testosterone (T) were determined by radioimmunoassay after administration.Results The linear range of morroniside,sweroside,paeoniflorin and loganin were 0.24-2.40 μg (r=0.999 6),0.14-1.38 μg(r =0.999 1),0.12-1.20 μg(r =0.999 1),0.24-2.40 μg(r =0.999 4) respectively.The recovery and relative standard deviation of them were (100.22 ± 1.80) ％,(100.83 ± 1.94) ％,(102.40 ± 1.47) ％,(101.40 ± 1.50) ％,and RSD were 1.80％,1.92％,1.44％,1.50％.The average content of morroniside,sweroside,paeoniflorin and loganin in Liuwei Dihuang Decoction water extract-alcohol soluble parts were calculate in terms of material were 0.20％,0.02％,0.06％,0.13％.The plasma level of cAMP in model group and experimental group were (8.20 ±0.63),(6.90 ±0.15) nmol · L-1 with significant different (P ＜0.01).The serum contents of FSH,E2,T in model group and experimental group were(0.54 ±0.10),(0.88 ±0.04)mU · mL-1;(13.93 ± 0.29),(15.48 ± 0.43) pg · mL-1;(2.23 ± 0.14),(5.63 ± 0.48) ng · mL-1 with significant different (P ＜ 0.01).Conclusion It is found that ethanol-soluble portion might be one of the bioactive component of Liuwei Dihaung Decoction to improve kidney Yin deficiency,and it mainly contains loganin,morroniside,sweroside and paeoniflorin.Its function mechanism of tonifying kidney may be related to the regulation of cAMP content and the levels of hormones in the HPG axis.