Mechanism analysis of repeatedly steamed and sundried Rehmanniae Radix Praeparata in delaying brain aging in ovariectomized mice based on proteomics / 中国中药杂志

The present study explored the effect and mechanism of repeatedly steamed and sundried Rehmanniae Radix Praeparata(RRP) in delaying brain aging in ovariectomized mice. After ovariectomy, the mice were randomly divided into a model group, an estradiol valerate group(0.3 mg·kg~(-1)), and low-(1.0 g·kg~(-1)), medium-(2.0 g·kg~(-1)), and high-dose(4.0 g·kg~(-1)) RRP groups, and a sham operation group was also set up, with 15 mice in each group. One week after the operation, intragastric administration was carried out for 15 consecutive weeks. The step-down test and Morris water maze test were used to detect the behavioral changes of mice. HE staining and Nissl staining were used to observe the morphological changes of mouse brain tissues. Immunohistochemistry was used to detect the expression of Aβ and ER_β in mouse brain tissues. The serum estrogen levels and cholinesterase and cholinesterase transferase levels in brain tissues of mice were detected by assay kits. The extracted hippocampal protein was detected by the Nano-ESI-LC-MS system, identified by the Protein Discovery, and analyzed quantitatively and qualitatively by the SIEVE. The PANTHER Classification System was used for GO analysis and KEGG pathway enrichment analysis of the differential proteins. Compared with the sham operation group, the model group showed decreased learning and memory ability, shortened step-down latency(P<0.05), prolonged escape latency(P<0.05), reduced platform crossings and residence time in the target quadrant, scattered nerve cells in the hippocampus with enlarged intercellular space, increased expression of Aβ-positive cells(P<0.05), declining expression of ER_β-positive cells and estrogen level(P<0.05), and weakened cholinergic function(P<0.05). Compared with the model group, the RRP groups showed improved learning and memory ability, prolonged step-down latency(P<0.05), increased estrogen level(P<0.05), neatly arranged nerve cells in the hippocampus with complete morphology, declining Aβ-positive cells, and elevated expression of ER_β-positive cells. A total of 146 differential proteins were screened out by proteomics, and KEGG pathway enrichment yielded 75 signaling pathways. The number of proteins involved in the dopaminergic synapse signaling pathway was the largest, with 13 proteins involved. In summary, RRP can delay brain aging presumedly by increasing the level of estrogen, mediating the dopaminergic synapse signaling pathway, and improving cholinergic function.

Relationship between Muscle Mass of Limbs and aGVHD in Patients with Allogeneic Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

OBJECTIVE@#To explore the correlation of limb muscle mass and acute graft-versus-host disease.@*METHODS@#Clinical data from 144 patients treated by allo-HSCT in Guangzhou First People's Hospital were collected and analyzed retrospectively. The age, sex, diagnosis, donor age, sex of the donors, preparative regimen, ATG dose, HLA match, graft source, and number of infused stem cells of the patients were collected as baseline information. Meanwhile, bioelectrical impedance principle (BIA) was used to measure the limb muscle mass, body weight, body mass index (BMI), waist-to-hip ratio, upper arm muscle circumference, triceps skinfold thickness, and body fat rate of the patients before and after transplantation, so as to compare the changes of limb muscle mass and investigate its correlation with aGVHD.@*RESULTS@#It was found that 61.11% of allo-HSCT patients showed muscle mass loss, and the proportion of male and female was 35.42% and 25.69%, respectively. There were reduction in the body weight, BMI, upper arm muscle circumference and muscle mass of limbs after transplantation as compared with those before transplantation (P<0.05). By comparing with the cumulative incidence of aGVHD between the patients in low muscle mass group and normal muscle mass group, it was found that the cumulative incidence of Ⅱ-Ⅳdegree aGVHD in patients with low muscle mass (30.38%) was higher than those with normal muscle mass (8.93%), which showed statistical difference (P<0.05). Univariate analysis showed that muscle mass, the sex of the donors, and preparative regimen were the influencing factors of aGVHD (P<0.05). Binary logistic regression showed that low muscle mass was the independent risk factor affecting aGVHD (P<0.05).@*CONCLUSION@#Patients treated by allo-HSCT shows a decline in muscle mass after transplantation, and the incidence of aGVHD is high in patients with low muscle mass. Therefore, the assessment of muscle quality in early stage in patients with HSCT can facilitate earlier detection of aGVHD.

Anti-leukemia effect of oridonin on Ph(+) acute lymphoblastic leukemia cell SUP-B15 / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the anti-leukemia effect of oridonin on Ph(+) acute lymphoblastic leukemia (ALL) cell line SUP-B15.</p><p><b>METHODS</b>Human Ph(+) ALL cell line was cultured in vitro. The 50% inhibition concentration (IC(50)) of oridonin against SUP-B15 cell line was examined using modified MTT assay. The cellular morphologic changes were observed using a light microscope. The percent of apoptosis of SUP-B15 cell line after drug treatment was evaluated by flow cytometric analysis. The active levels of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK, STAT5 signaling pathways and the expression levels of Bcl-2 and BAX were examined by Western blot.</p><p><b>RESULTS</b>Oridonin inhibited the growth of SUP-B15 cell line in both time- and dose-dependent manner with the IC(50) of oridonin as (7.08 ± 1.21) µmol/L after 72 h treatment. The cellular membrane of SUP-B15 cell line treated with oridonin became unsharp, some of them disintegrated. Oridonin induced apoptosis in SUP-B15 cell line with the apoptosis rates following 0, 5, 10 µmol/L oridonin treatment for 24 h were (6.67 ± 0.83)%, (18.30 ± 1.79)% and (37.63 ± 7.12)%, respectively. Oridonin inhibited activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STAT5 signaling pathways, which were constitutively activated in SUP-B15 cell line, down-regulated the level of anti- apoptotic protein Bcl-2 and up-regulated the expression of pro-apoptotic protein Bax.</p><p><b>CONCLUSION</b>Oridonin exerted anti-leukemia effect in Ph(+)ALL cell line SUP-B15 by inhibiting the activation of ABL kinase and its downstream Akt/mTOR, Raf/MEK/ERK and STAT5 signaling pathways, down-regulating the expression of Bcl-2 and up-regulating the expression of BAX.</p>

The effect of knockdown of transcription factor SCL/TAL-1 gene on the erythroid differentiation in EPO-induced K562 cell line / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of transcript factor SCL/TAL-1 gene in the erythroid differentiation through the knockdown of SCL/TAL-1 mRNA by RNA interference.</p><p><b>METHODS</b>The plasmid of pTRIP-dU3-RNAiTALh-EF1a-GFP with SCL/TAL1 shRNA was transfected into EPO-induced K562 cell line with erythroid differentiation via lentiviral vector system and the expression of SCL/TAL-1 mRNA decreased. The plasmid pTRIP-dU3- RNAiluc-EF1-GFP expressing EGFP gene was as control. The mRNA levels of SCL/TAL-1 and erythroid related RhD, GPA, CD47 in the cell lines were detected by RT-PCR, and erythroid antigen CD71, CD235a were examined by flow cytometry.</p><p><b>RESULTS</b>(1) After 48 h of transfect, more than 95% of K562 cells were GFP positive, indicating the infection rate of the plasmids in the K562 cells more than 95%. (2) The results of RT-PCR showed SCL/TAL-1 mRNA expression in the K562 cell line of knockdown of SCL/TAL-1 was significantly lower than that in the control (P < 0.05). The mRNA levels of CD47 and RhD were also significantly lower, however, GPA decreased slightly in comparison with the control. (3) The expressions of CD71 and CD235a markedly reduced in the K562 cell line of knockdown of SCL/TAL-1 with positive rates as 10.4% and 76.5%, while the positive rates in the control as 94.3% and 83.6%.</p><p><b>CONCLUSION</b>Our findings suggested that transcription factor SCL/TAL-1 might play an positive role in erythroid differentiation.</p>

Correlation between postmortem intervals and the changes of electrical conductivity in vitreous humor in rabbits after death / 法医学杂志

OBJECTIVE@#To investigate the correlation between postmortem interval (PMI) and the changes of electrical conductivity in vitreous humor in rabbits after death.@*METHODS@#The changes of electrical conductivity in vitreous humor in rabbits were measured using the conductivity meter under 30 degrees C during 0-48 hours and 20 degrees C during 0-120 hours after death.@*RESULTS@#Electrical conductivity in vitreous humor in rabbits increased gradually under 30 degrees C and 20 degrees C from 0 to 48 hours and from 0-120 hours after death. The formulae of the relationship between PMI and conductivity under 30 degrees C and 20 degrees C were obtained by statistical analysis and the correlation coefficients were 0.970 and 0.983 (both P < 0.01), respectively.@*CONCLUSION@#The increase of electrical conductivity in vitreous humor in rabbits after death may be used as the relatively objective parameter for PMI estimation.

In vitro study on microleakage of three adhesive materials / 上海第二医科大学学报

Objective To evaluate the influence of self-etch adhesive,total-etch adhesive and glass ionomer cement on the marginal microleakage of class II restorations. Methods Thirty human premolars were randomly divided into 3 groups(n=10),and cuboid class II cavities(4.0 mm?3.5 mm?2.5 mm) were prepared.Restoration was performed using self-etch adhesive+nano-resin(self-etch group),total-etch adhesive + nano-resin(total-etch group) or glass ionomer cement(glass ionomer group).Half of each group underwent 200 thermocyclings and the other half underwent 500 thermocyclings.The specimens were immersed in 0.5% basic fuchsin for staining.Each tooth was then evaluated the microleakage at the axial wall and the gingival wall section by section under a stereomicroscope.The data were statistically analyzed. ResultsSelf-etch group had significantly more miroleakage than total-etch group and glass ionomer group after 200 and 500 thermocyclings(P