Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.

Total flavonoids of Dracocephalum moldavica L. (TFDM) is an effective component extracted and isolated from the traditional Uighur medicinal herb Cymbidium fragrans. Cymbidium fragrans has the effects of tonifying the heart and brain, promoting blood circulation and resolving blood stasis, and has been widely used in the treatment of cardiovascular and cerebrovascular diseases for a long time. The purpose of this study was to determine the effect of total flavonoids from Cymbidium fragrans on hypoxia/re-oxygenation (H/R) injury in H9c2 (rat cardiomyocytes) cells and its mechanism. A model (H/R) of hypoxia/re-oxygenation injury in H9c2 cells was established using hypoxia and glucose deprivation for 9 h combined with re-oxygenation and rehydration for 2 h to simulate myocardial ischemia-reperfusion injury. The effects of total flavonoids from Cymbidium fragrans on cell viability, markers of myocardial cell damage, oxidative stress levels, and reactive oxygen radical (ROS) content were investigated, Western blot was used to detect the expression of vascular endothelial growth factor B (VEGF-B) and adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) pathway related proteins. The results showed that the total flavonoids of Cymbidium fragrans significantly increased the viability of myocardial cells after H/R injury, and decreased the content of lactate dehydrogenase (LDH) and creatine kinase isozyme (CK-MB) in the cell supernatant. It significantly reduced malondialdehyde (MDA), increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and decreased intracellular ROS and nitric oxide (NO) content. Western blot analysis showed that the total flavonoids of Cymbidium fragrans decreased Bax levels in H9c2 cells damaged by H/R and increased Bcl-2 expression. Total flavones of Cymbidium fragrans upregulate VEGF-B/AMPK pathway related proteins VEGF-B, vascular endothelial growth factor receptor 1 (VEGFR-1), neuropilin 1 (NRP-1), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), phosphorylated adenosine monophosphate activated protein (p-AMPK) and phospho mechanistic target of rapamycin (p-MTOR) levels. The above research results indicate that the total flavonoids of Cymbidium can significantly reduce the H/R injury of myocardial cells, which may be related to the upregulation of VEGF-B/AMPK pathway and inhibition of oxidative stress response.

Hyperlipidemia,as one of the severe risk factors of cardiovascular disease,could easily trigger atherosclerosis,coronary heart disease,peripheral vascular disease,pancreatitis,etc.,and could also increase the incidence of type 2 diabetes and fatty liver disease. Improving dyslipidemia could slow down the progression of atherosclerosis and reduce the risk of coronary heart disease. This is of great importance for prevention and treatment of cardiovascular disease. Phytosterols are natural active ingredients in plants. Many researches have shown that phytosterols have significant lipid-lowering activity,which could effectively lower blood cholesterol and triglyceride levels. Foods containing phytosterols have been widely used as therapeutic diets for improving dyslipidemia. In the early years,it was believed that the lipid-lowering effect of phytosterols was achieved by competitively inhibiting the absorption of dietary cholesterol in the intestine since phytosterols had similar chemical structures with cholesterol. In further researches in recent years,more progress has been made in the lipid-lowering mechanisms of phytosterols. In this paper,PubMed and Web of Science were used to review the cholesterol-lowering and triglyceride-lowering mechanisms of phytosterols according to the available data published,so as to use phytosterols more rationally in clinical application to improve hyperlipidemia and other induced diseases.
Cholesterol ; Diabetes Mellitus, Type 2 ; Humans ; Hyperlipidemias ; Hypolipidemic Agents/pharmacology* ; Phytosterols/pharmacology* ; Triglycerides

AIM:To investigate the synergistic effect of imperatorin on enhancing the anti -tumor effect of TNF-related apoptosis-inducing ligand(TRAIL)on breast cancer and the mechanisms.METHODS:T-47D and MCF-7 breast cancer cells were divided into control group ,imperatorin group,TRAIL group,imperatorin+TRAIL group and imperatorin+TRAIL+death receptor 5(DR5)siRNA group.The viability of T-47D and MCF-7 cells was measured by MTT assay. The apoptosis and mitochondrial membrane potential in T-47D cells were analyzed by flow cytometry.Western blot and flow cytometry analysis were performed to evaluate the expression of DR 5 on T-47D cell surface and the activation of caspase-8 and caspase-3.RESULTS:Imperatorin significantly enhanced the inhibition of cell viability induced by TRAIL of T -47D and MCF-7 cells,and significantly increased the apoptosis of T-47D cells induced by TRAIL.Imperatorin treatment ob-viously induced upregulation of DR5 expression and production of reactive oxygen species in the T-47D cells.In addition,imperatorin enhanced the TRAIL-induced damage of mitochondrial membrane potential and activation of caspase -8 and caspase-3.CONCLUSION:Imperatorin enhances the anti-tumor effect of TRAIL on breast cancer via upregulating the ex-pression of DR5.

Biomarkers ; blood ; Cell-Derived Microparticles ; metabolism ; Coronary Disease ; blood ; Humans ; Inflammation ; blood

OBJECTIVETo isolate, identify and culture human spermatogonial stem cells (SSC) and then obtain purified and enriched human SSCs for research and application.

METHODSWe detected the expression of CD90 in the human testis using the immunofluorescence technique and isolated human testicular spermatogenic cells by two-step enzymatic digestion, followed by differential plating and magnetic-activated cell sorting (MACS) with CD90 as an SSC marker. Then we identified the isolated CD90-positive spermatogenic cells by RT-PCR and immunocytochemistry, and meanwhile cocultured them with Sertoli cells in SG medium in vitro.

RESULTSThe isolated CD90-positive cells showed a relatively homogeneous characteristic in size and morphology and expressed the genes specific for human SSCs, with high expressions (90.5%) of GFRA1, GPR125, and UCHL1. After coculture with Sertoli cells in the SG medium for 2 weeks, the isolated CD90-positive cells maintained a good activity.

CONCLUSIONCD90 can be regarded as a speci- fic marker for human SSCs and used to obtain highly enriched human SSCs by differential plating and MACS. Furthermore, the isolated human SSCs can be cultured in SG medium in vitro.

Adult Stem Cells ; cytology ; Biomarkers ; metabolism ; Cell Separation ; methods ; Cell Shape ; Cell Size ; Coculture Techniques ; Glial Cell Line-Derived Neurotrophic Factor Receptors ; metabolism ; Humans ; Immunohistochemistry ; Male ; Receptors, G-Protein-Coupled ; metabolism ; Sertoli Cells ; Spermatogonia ; cytology ; Testis ; metabolism ; Thy-1 Antigens ; isolation & purification ; metabolism ; Ubiquitin Thiolesterase ; metabolism

This study was purposed to investigate the protective effects of lipoprotein HS-6101(6101) on rhesus monkey total body irradiated with 7.0 Gy ⁶⁰Coγ-ray. A total of 30 health adult rhesus monkeys were randomly divided into symptomatic therapy (ST), WR2721 and HS-6101 30, 90 and 270 mg/kg groups (n = 6), the rhesus monkeys of each groups were injected with physiological saline 0.3 ml/kg, WR-2721 30 mg/kg, or HS-6101 30, 90 and 270 µg/kg, respectively. All agents were once intramuscularly injected at 1 hr prior irradiation. General observation, peripheral blood cell counts, colony forming unite assay of bone marrow hemopoietic progenitor cells, and histopathological examination were performed. The results showed that animals in symptomatic therapy group begin to die on the 13(th) day and 4 animals died within 24 days, the average survival time was 18.2 ± 4.3 days; 2 animals in WR-2717 groups died on day 15.8 and day 18.5 post irradiation respectively. 1 animal in HS-6101 270 mg/kg group died on day 35.8, all other animals survived. Nadirs of peripheral blood white blood cells, neutrophils and platelets of animals in HS-6101 treatment groups were significantly higher than those in other 2 groups including ST and WR-2721 groups, and the hemopoietic recovery were also significantly speeding up(P < 0.05 and 0.01). In vitro results showed that HS-6101 obviously promoted 7.0 Gy ⁶⁰Coγ irradiated monkey's bone marrow mononuclear cells to form various hematopoietic progenitor cell colonies (P < 0.05 and 0.01) . Compared with symptomatic therapy and WR-2717 groups, bone marrow histopathological changes in HS-6101 treatment groups showed more active hemopoietic cell proliferation and higher density structure. It is concluded that HS-6101 90 µg/kg treatment can promote the bone marrow recovery of 7.0 Gy ⁶⁰Coγ irradiated monkey, alleviate their animal symptom, simplify the treatment measures and improve the animal survival rate. The HS-6101 shows remarkable radioprotective effects as compared with the currently internationally acknowledged radioprotectant of WR-2721.
Amifostine ; Animals ; Blood Cell Count ; Blood Platelets ; Bone Marrow ; Bone Marrow Cells ; Hematopoietic Stem Cells ; Hematopoietic System ; drug effects ; radiation effects ; Lipoproteins ; pharmacology ; Macaca mulatta ; Radiation Injuries ; drug therapy ; Survival Rate

This study was purposed to investigate the immune reconstitution of T-cells in patients who received haploidentical hematopoietic stem cell transplantation (hiHSCT). The peripheral blood was harvested from 22 patients before transplantation and at month 1, 3, 6 after hiHSCT. The proportions of T lymphocyte subtypes including CD3(+), CD4(+), CD8(+), CD45RO(+), and CD45RA(+)CD62L(+) were analyzed by flow cytometry, followed by the calculation of T cell numbers according to the amounts of peripheral blood leukocytes. Adenosine triphosphate (ATP) value in CD4(+) T cells was measured by ImmuKnow method to evaluate the function of lymphocytes. The results showed that the CD3(+) cell absolute value before transplantation was 833.75 ± 359.84/µl, but those values at month 1, 3, 6 after transplantation were 318.87 ± 266.71/µl, 1006.76 ± 512.32/µl and 1296.38 ± 958.77/µl respectively. The CD4(+) cell absolute value before transplantation was 336.99 ± 211.11/µl, but such values at month 1, 3, 6 after transplantation were 45.89 ± 44.21/µl, 142.97 ± 114.85/µl, and 181.78 ± 120.61/µl respectively. The CD8(+) cell absolute value before transplantation was 430.21 ± 159.48/µl, but those values at month 1, 3, 6 after transplantation were 230.44 ± 195.89/µl, 621.64 ± 318.83/µl, and 823.07 ± 633.55/µl respectively. The CD4(+)CD45RO(+) memory T cell absolute value before transplantation was 227.44 ± 73.34/µl, but such values at month 1, 3, 6 after transplantation were 43.47 ± 43.40/µl, 138.69 ± 110.17/µl, 147.73 ± 82.94/µl respectively. The CD8(+)CD45RO(+) memory T cell absolute value before transplantation was 212.70 ± 98.48/µl, but such values at month 1, 3, 6 after transplantation were 184.76 ± 168.65/µl, 445.90 ± 252.50/µl, 519.80 ± 475.53/µl respectively. CD4(+)CD45RA(+)CD62L(+) naive T cell number before transplantation was 68.94 ± 59.74/µl, but such cell numbers at month 1, 3, 6 after transplantation decreased to 2.44 ± 2.93/µl, 3.14 ± 3.48/µl, 23.22 ± 38.38/µl respectively. The CD8(+)CD45RA(+)CD62L(+) naive T cell absolute value before transplantation was 124.82 ± 60.95/µl, but those values at month 1, 3, 6 decreased to 19.37 ± 17.71/µl, 76.63 ± 50.85/µl, and 114.49 ± 174.29/µl respectively. The ATP value in CD4(+) T cells decreased to 210.19 ± 119.37 ng/ml at month 1 after transplantation and increased to 280.62 ± 110.03 ng/ml at month 3, and 357.28 ± 76.18 ng/ml at month 6 after transplantation. It is concluded that CD8(+) memory T cell reconstruction contributes critically to T cell recovery early after hiHSCT, while the thymic output function remains low. However, T cell function recovers to normal range at month 3 after transplantation.
Adolescent ; Adult ; CD8-Positive T-Lymphocytes ; cytology ; Child ; Child, Preschool ; Female ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Killer Cells, Natural ; immunology ; Lymphocyte Count ; Male ; T-Lymphocyte Subsets ; immunology ; Young Adult

OBJECTIVETo detect the changes of cardiovascular system at different altitudes, so as to prevent and predict the susceptibility to acute mountain sickness.

METHODSThe test was performed with noninvasive cardiovascular monitor and electrocardiogram in healthy male Chinese members of the 25th and 26th expeditions to the antarctic kunlun station at different altitudes (40 m, 3 650 m and 4 300 m).

RESULTSCompared with 40 m, from 3 650 m to 4 300 m, heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), systemic vascular resistance (SVR), systemic vascular resistance index (SVRI) significantly increased (P < 0.05); cardiac output (CO), cardiac index (CI), stroke volume (SV), stroke index (SI), velocity index (VI), acceleration index (ACI), left ventricular ejection time (LVET) significantly decreased (P < 0.05) and pre-ejection period(PEP) decreased with no significance (P > 0.05).

CONCLUSIONSVR significantly increased but contractile and blood-pumping function of left ventricular decreased inversely associated with the Q-Tc interval, as the altitude is increasing.

Adult ; Altitude ; Cardiac Output ; Electrocardiography ; Heart Rate ; Hemodynamics ; Humans ; Hypoxia ; Male

Objective To observe expression of neuron specific enolase (NSE) gene in the brain of rats induced by iodine excess. Methods One hundred and fifty one-month weaning Wistar rats were divided into ten groups according to 5 × 2 factorial experiment. Rats were fed with normal feedstuff and water of a series of iodine concentration by adding potassium iodide respectively: norrmal iodine (NI), five-fold high iodine (5HI), ten-fold high iodine(10HI), fifty-fold high iodine(50HI) and one hundred-fold iodine(100HI). After these rats were fed for three or six months, rat serum thyroid hormones were measured by radioimmunoassay including TT4, TT3, FT4, FT3, rT3 and the mRNA level of NSE in rat brain tissue was studied using RT-PCR technique. Results The levels of serum TT4 and TT3 were significantly different in five iodine level groups(F values were 18.867,27.287, both P < 0.01). The interaction between time and iodine level in TT4 was significant in our study(F values were 2.486, P < 0.05). The levels of TT4 and TT3 of 100 HI group at third and sixth month were lower than those of NI, 5HI, 10HI, 50HI groups in the same period (all P < 0.01). The levels of serum FT4, FT3 and rT3 were significantly different at different time(F values were 4.968,27.046,59.776 respectively, P < 0.05 or < 0.01) and in different iodine level groups(F values were 33.058,28.420,17.482 respectively, all P < 0.01). Moreover, the interaction between time and iodine level in FT3 and rT3 was significant in our study(F values were 6.894,5.233 respectively, both P < 0.01). FT4, FT3 and rT3 in 100HI group were lower than that of other iodine dosage groups at the same time (P < 0.05 or < 0.01). The levels of NSE mRNA in brain tissue was significantly different in five iodine level groups (F values were 29.006, P < 0.05). The levels of mRNA NSE of 100HI group in both three and six months (0.61 ± 0.19,0.61 ± 0.22) were all lower than that of any other groups[NI(0.73±0.13 and 0.72 ±0.26), 5HI (0.72 ± 0.15 and 0.72±0.16), 10HI (0.73 ±0.32 and 0.70±0.13), 50HI(0.71±0.18 and 0.69±0.31), all P < 0.05]. The results of correlation analyses show that the levels of serum FT3 and FT4 had correlations with the levels of NSE mRNA (P < 0.05) both in three and six months(r values were 0.987, 0.969 in three month, and 0.890, 0.910 in six month respectively). Conclusions The expression of NSE gene can tolerant the excess of iodine to a certain extent. Exposure to heavy excess iodine(100HI) can decrease the mRNA level of NSE gene. FT4 and FT3 may both have important roles on the regulation of NSE mRNA induced by excess iodine.