Histone Variant H2A.Z Enhances the Stability of Nucleosomes under the Effect of Salt Ions / 中国生物化学与分子生物学报

In eukaryotes, the basic structural unit of chromatin is the nucleosome, and genomic DNA iscompressed in chromatin. The presence of nucleosomes usually inhibits the biological processes that occuron the DNA templates, such as transcription, replication, repair, and recombination,. The histonevariant H2A.Z can regulate the structure of chromatin and affect the transcription process of genes, but itsdetailed regulation mechanism remains unclear. In this paper, the method of Förster resonance energytransfer (FRET) was used to detect the influence of sodium chloride, potassium chloride, manganesechloride, calcium chloride and magnesium chloride on the dissociation of nucleosomes. Then, the stabilitydifferences between the nucleosomes containing the histone variant H2A. Z and the conventionalnucleosomes were compared under the action of salt ions. Widom 601 DNA sequence was labeled withdual fluorescence Cy3 and Cy5, and the dissociation of nucleosomes was reflected by the change offluorescence signal value. FRET results showed that the dissociation speed of nucleosomes containing thehistone variant H2A. Z under the action of sodium chloride, potassium chloride, manganese chloride, calcium chloride and magnesium chloride is slower than that of canonical nucleosomes, and the influenceof calcium chloride, manganese chloride and magnesium chloride on dissociation is more obvious thansodium chloride, potassium chloride. The results of electrophoresis analysis showed that the dissociationrate of nucleosomes containing histone variant H2A. Z was significantly lower than that of canonicalnucleosomes at 75℃. Fluorescence thermal shift (FTS) was used to further analyze the stability ofnucleosomes containing histone variant H2A.Z. It was found that the fluorescence signals of the two typesof nucleosomes showed two distinct growth periods, and the fluorescence signals generated in thedissociation process of the two types of nucleosomes showed two distinct growth periods. The temperaturecorresponding to the first increasing period of fluorescence signal in the dissociation process of H2A. Z-containing nucleosome is significantly higher than that in the dissociation process of the canonicalnucleosome, which indicated that the dissociation and denaturation temperature of the H2A.Z / H2B dimerin the nucleosome is higher than that of the canonical H2A / H2B dimer, and the H2A. Z-containingnucleosomes have high thermal stability. The results indicated that the structure of nucleosomes containingthe histone variant H2A.Z is more stable than that of canonical nucleosomes.

Effect of Ranae Oviductus on Follicular Development Based on PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the effect of Ranae Oviductus (RO) on ovarian follicular development， phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway， and pregnancy function in rats, and the estrogen-like mechanism of OR. Method:Seventy female Wistar rats were randomly divided into a normal group， a progynova+ luteohormone group (1 mg·kg^-1+40 mg·kg^-1), a clomiphene group (10 mg·kg^-1), and high-dose(400 mg·kg^-1) and low-dose(200 mg·kg^-1) RO groups. Rats were administered correspondingly by gavage for eight weeks. After seven weeks of intragastric administration, the estrus cycle of all rats was measured. After eight weeks of intragastric administration， four rats from each group were selected to give birth. For other rats, blood was collected on the day of estrus， and the serum levels of estradiol (E₂)，progesterone (P)， testosterone (T)， follicle-stimulating hormone (FSH)， and luteotropic hormone (LH) were detected. Uterus and ovaries were extracted and weighed to calculate organ index. One ovary was made into pathological sections, and the follicles at different developmental stages and corpus luteum were counted. Real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）and Western blot were performed on the other ovary to detect mRNA and protein changes in the PI3K/Akt signaling pathway. Forty female Kunming mice were randomly divided into a normal group and RO groups (400 mg·kg^-1) with 14 days， 28 days， and 56 days of intervention. Mice in the RO groups were raised with male mice in cages after intragastric administration of OR for 14， 28， and 56 days， respectively. After 18 days, the number of intrauterine fetuses on both sides and the number of stunted fetuses were counted. Result:After eight weeks of intragastric administration of OR， the rats showed decreased uterine index (<italic>P</italic><0.05)， declining serum LH (<italic>P</italic><0.05)， reduced luteum (<italic>P</italic><0.01)， dwindled primary follicles (<italic>P</italic><0.05)， and increased rate of follicle atresia (<italic>P</italic><0.01). Additionally， more luteal or interstitial glands degenerated into interstitial structures in the ovarian cortex in a short time. The mRNA expression levels of PI3K and Akt in the ovary were elevated (<italic>P</italic><0.01)， while the mRNA expression levels of mTOR and PTEN were reduced (<italic>P</italic><0.01). The phosphorylation level of Akt protein showed a downward trend without significant difference. For the rats， the number of fetuses was decreased (<italic>P</italic><0.05). The pregnancy rate of mice was decreased to varying degrees after administration of RO for different durations， with the lowest in the 14 day RO group， as low as 30%. After 28 days of intragastric administration of RO， the difference in left and right uterine pregnancy increased (<italic>P</italic><0.05). Conclusion:Long-term administration of RO can lead to premature ovarian failure by over-stimulating the ovary， which is similar to clomiphene. Short-term administration can result in decreased pregnancy rate， excessive ovulation on one side， and inhibition of ovulation on the other side. The influence on follicles needs further exploration.

Analysis of Health Service Utilization and its Influencing Factors among Patients with Pneumoconiosis in China / 生物医学与环境科学（英文）

Pneumoconiosis, an interstitial lung disease that occurs from breathing in certain kinds of damaging dust particles, is a major occupational disease in China. Patients diagnosed with occupational pneumoconiosis can avail of free medical treatment, whereas patients without a diagnosis of occupational diseases cannot not claim free medical treatment in most provinces from the government before 2019. This study aimed to analyze the priority of medical facility selection and its influencing factors among patients with pneumoconiosis. A total of 1,037 patients with pneumoconiosis from nine provinces in China were investigated. The health service institutions most frequently selected by the patients were county-level hospitals (37.5%). The main reason for the choice was these hospitals' close distance to the patients' homes (47.3%). The factors for the choice of health care institutions were living in the eastern region (

Effectiveness of functional training and diet intervention on functional movement and physical health of obese male college students / 中国学校卫生

Objective@#To reveal the effect of functional training & diet intervention on functional movement screen (FMS) and physical health of obese college male students.@*Methods@#Totally 56 obese college male students were divided into experimental group and control group on average and the experimental group took functional training & diet intervention, 2 times per week, with a total of 15 weeks. The control group was only given routine physical exercise without diet intervention. Indexes including seven FMS indicators, seven body composition indicators and five physical fitness indicators were monitored before and after experiment.@*Results@#The results showed that except for shoulder flexibility, other FMS indexes (vault, hurdle, straight lunge, trunk stability and trunk rotation stability) of experimental group significantly increased after functional training & diet intervention(t=-2.11, -2.27, -2.22, -2.08, -2.21, P<0.05). The total score of FMS increased significantly from 12.07 to 14.06(t=-4.22, P<0.05). The body composition index including fat content, body water, body fat ratio and BMI decreased to 23.97 kg, 38.83 kg, 26.34% and 27.79 kg/m2(t/χ2=-5.85, -8.30, 9.05, 7.23, P<0.05). Protein increased significantly to 19.00 kg(t=-5.75, P<0.05). The comprehensive score of body composition in the experimental group increased significantly from 64.00 to 66.71(t=-6.27, P<0.05). The scores of experimental group physical fitness indicators including 50 m, sit forward, standing long jump, pull up and 1 000 m increased to 8.24 s, 14.45 cm, 206.61 cm, 3.07 and 278.27 s(t=5.05, -3.96, -7.46, -3.62, 4.40, P<0.05).@*Conclusion@#The study indicates that 15 weeks of physical function training & dietary intervention could effectively improve the body shape, body composition and motor dysfunction of obese male college students, thus to reduce sports risk and improve physical health.

Enhanced Tendon-to-Bone Healing of Chronic Rotator Cuff Tears by Bone Marrow Aspirate Concentrate in a Rabbit Model

BACKGROUND: To evaluate the influence of bone marrow aspirate concentrate (BMAC) on tendon-to-bone healing in a rabbit rotator cuff model and to characterize the composition of growth factors in BMAC. METHODS: In this in vivo study, 40 rabbits were allocated into five groups: control (C), repair + saline (RS), repair + platelet-rich plasma (PRP; RP), repair + BMAC (RB) and repair + PRP + BMAC (RPB). A tear model was created by supraspinatus tendon transection at the footprint. Six weeks after transection, the torn tendon was repaired along with BMAC or PRP administration. Six weeks after repair, shoulder samples were harvested for biomechanical and histological testing. Ten rabbits were used for processing PRP and BMAC, followed by analysis of blood cell composition and the levels of growth factors in vitro. RESULTS: The ultimate load-to-failure was significantly higher in RPB group compared to RS group (p = 0.025). BMAC-treated groups showed higher values of biomechanical properties than RS group. The histology of BMAC-treated samples showed better collagen fiber continuity and orientation than RS group. BMAC contained significantly higher levels of the several growth factors than PRP. CONCLUSIONS: Locally administered BMAC enhanced tendon-to-bone healing and has potential for clinical applications.

IL-32 mRNA Expression of Bone Marrow Stromal Cells and Its Correlation with Cell Apoptosis in Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the IL-32 mRNA expression of bone marrow stromal cells and its correlation with apoptosis of bone marrow mononuclear cells in patients with myelodysplastic syndrome (MDS).</p><p><b>METHODS</b>Bone marrow samples from 26 MDS patients and 10 iron deficiency anemia (IDA, as control) patients were collected, RT-PCR was used to detect the IL-32 mRNA expression of bone marrow stromal cells, and the apoptosis of bone marrow mononuclear cells was detected by flow cytometry with Annexin V-FITC/PI dowble staining. The born marrow lymphocytes and NK cells were detected by means of direct immunofluorescence labeling whole blood hemolysis and flow cytometry.</p><p><b>RESULTS</b>IL-32 mRNA expression of bone marrow stromal cells in the MDS patients was significantly higher than that of control group, the IL-32 mRNA expression of bone marrow stromal cells in patients with RA, RAS and RCMD was significantly higher than that in patients with RAEB. There was no obvious difference between RAEB and the control groups. The apoptosis of bone marrow mononuclear cells in MDS group was significantly higher than that in the control group, the apoptosis of bone marrow mononuclear cells in patients with RA, RAS and RCMD was significantly higher than that in RAEB. There was no significant difference between RAEB group and control group. The IL-32 mRNA expression in bone marrow stromal cells significantly correlated with the apoptosis of bone marrow mononuclear cells in MDS patients. The NK cell number in born marrow of MDS patients and the control group had no significant difference.</p><p><b>CONCLUSION</b>The expression of IL-32 mRNA in bone marrow stromal cells significantly relates with the apoptosis of MDS cells, and the secretion of IL-32 by bone marrow stromal cells may be one of the reasons for the apoptosis of MDS bone marrow cells. It is speculated that the abnormal MDS bone marrow microenvironment is involved in the apoptosis of bone marrow cells.</p>

Mutation detection of mitochondrial DNA D-loop region in bone marrow cells of acute leukemia / 中国实验血液学杂志

This study was aimed to detect the mutations and microsatellite instability (mtMSI) in mitochondrial DNA (mtDNA) D-loop region in bone marrow cells of acute leukemia (AL) patients, and to analyze their relationship with the pathogenesis of AL. 19 cases of newly diagnosed AL were enrolled in this study. Through extracting mtDNA, the D-loop region was amplified by polymerase chain reaction (PCR), the sequences of PCR products were detected by the pros- and cons-direct sequencing methods. The sequencing results were compared with the revised Cambridge reference sequence (rCRS) and the relevant database (MITOMAP database, GenBank database, mtDB database). The results showed that the mutation rate of mtDNA D-loop region in AL was 79% (15/19). 215 variations (35 mutations, 180 SNP) and a kind of mtMSI in the D-loop region were detected. A new type of mutation nt150 C-CT was found. Also, there was no significant difference in the number of mutations between patients with different ages and different types of AL (AML, B-ALL). It is concluded that there is high frequency of mutations in the mtDNA D-loop, and the mutations may be associated with the pathogenesis of AL.

Challenges and strategies of drug innovation / 药学学报

Drug research involves scientific discovery, technological inventions and product development. This multiple dimensional effort embodies both high risk and high reward and is considered one of the most complicated human activities. Prior to the initiation of a program, an in-depth analysis of "what to do" and "how to do it" must be conducted. On the macro level, market prospects, capital required, risk assessment, necessary human resources, etc. need to be evaluated critically. For execution, drug candidates need to be optimized in multiple properties such as potency, selectivity, pharmacokinetics, safety, formulation, etc., all with the constraint of finite amount of time and resources, to maximize the probability of success in clinical development. Drug discovery is enormously complicated, both in terms of technological innovation and organizing capital and other resources. A deep understanding of the complexity of drug research and our competitive edge is critical for success. Our unique government-enterprise-academia system represents a distinct advantage. As a new player, we have not heavily invested in any particular discovery paradigm, which allows us to select the optimal approach with little organizational burden. Virtue R&D model using CROs has gained momentum lately and China is a global leader in CRO market. Essentially all technological support for drug discovery can be found in China, which greatly enables domestic R&D efforts. The information technology revolution ensures the globalization of drug discovery knowledge, which has bridged much of the gap between China and the developed countries. The blockbuster model and the target-centric drug discovery paradigm have overlooked the research in several important fields such as injectable drugs, orphan drugs, and following high quality therapeutic leads, etc. Prejudice against covalent ligands, prodrugs, nondrug-like ligands can also be taken advantage of to find novel medicines. This article will discuss the current challenges and future opportunities for drug innovation in China.

Globular adiponectin protects human umbilical vein endothelial cells against apoptosis through adiponectin receptor 1/adenosine monophosphate-activated protein kinase pathway / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Endothelial dysfunction is a key event in the onset and progression of atherosclerosis in diabetic patients. Apoptosis may lead to endothelial dysfunction and contribute to vascular complications. However, no study has addressed apoptosis in human umbilical vein endothelial cells (HUVECs) induced by an intermittent high-glucose media and its association with adiponectin receptor 1 (adipoR1), adipoR2, or adenosine monophosphate (AMP)-activated protein kinase (AMPK).</p><p><b>METHODS</b>HUVECs were cultured in continuous normal glucose (5.5 mmol/L), continuous high glucose (25 mmol/L), alternating normal and high glucose and mannitol. In the alternating normal and high-glucose media, HUVECs were treated under different conditions. First, cells were transfected with the adipoR1-specific small-interfering RNA (siRNA) and then stimulated with globular adiponectin (gAD). Second, cells were cultured in both gAD and the AMPK activator 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR). Third, cells were cultured in the AMPK inhibitor adenine-9-β-D-arabino-furanoside (araA), gAD, and in AICAR.</p><p><b>RESULTS</b>HUVEC apoptosis increased more significantly in an intermittent high-glucose medium than in a constant high-glucose medium. HUVEC apoptosis induced by an intermittent high-glucose medium was inhibited when the cells were pretreated with 3 µg/ml gAD, which rapidly activated AMPK and adipoR1 in HUVECs. However, adipoR2 was not activated.</p><p><b>CONCLUSIONS</b>We found that adipoR1, not adipoR2, is involved in mediating intermittent high-concentration glucose-evoked apoptosis in endothelial cells. gAD activated AMPK through adipoR1, leads to the partial inhibition of HUVEC apoptosis. A fluctuating glucose medium is more harmful than a constant high-glucose medium to endothelial cells.</p>

Survival analysis on advanced non-small cell lung cancer with a Buckley-James model / 中华流行病学杂志

Objective To analyze the risk factors related to survival time of advanced nonsmall cell lung cancer(NSCLC)and to establish a prediction model on survival time. Methods From 2004-2006, 184 patients with advanced NSCLC were enrolled in the Affiliated Hospital to the Nantong Midical College. Related risk factors were analyzed, using the Buckley-James model. Both actual and predicted survival time were compared by log-rank test. Results Through Buckley-James model analysis, data showed that KPS, clinical stage, treatment and pre-treatment hemoglobin were main influencing factors on survival time. Regression equation appeared to be lnMONTH=0.0108 KPS+0.0238 HB+0.4614 Ⅲb+0.8027 Ⅲa+0.3869(radiotherapy+chemotherapy)+0.507 (radiotherapy + operation)+ 0.6082(chemotherapy + operation)- 2.098. There was no statistical difference between the prediction and the actual models of survival time by log-rank test(P=0.575＞0.05). Conclusion KPS, clinical stage, treatment and pre-treatment hemoglobin might be associated. Both the prognosis of patients with advanced NSCLC and the prediction model seemed to have practical significances.

Expression of pituitary tumor transforming gene and C-myc gene in patients with multiple myeloma / 中国实验血液学杂志

The study was purposed to explore the expressions of pituitary tumor transforming gene and c-myc gene in patients with multiple myeloma (MM) and its relationship with pathogenesis of MM. Expressions of pituitary tumor transforming gene and c-myc gene mRNA in BMMNC from 33 patients with MM and 10 normal controls were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that the expressions of pituitary tumor transforming gene and c-myc gene mRNA were significantly higher in MM patients those that in normal controls (p<0.05). The expression of pituitary tumor transforming gene mRNA was significantly correlated with the expression of c-myc gene (r=0.801, p<0.05). In conclusion, the overexpressions of pituitary tumor transforming gene and c-myc gene may be related to the pathogenesis and progression of MM.

Effect of aspirin on high glucose-induced senescence of endothelial cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Endothelial cell senescence is accelerated under high glucose condition, which may contribute to the vascular complications in the diabetics. It has been proved that aspirin has multiple cytoprotective effects. This study aimed to investigate the effect of aspirin on high glucose-induced endothelial cell senescence and its possible mechanism.</p><p><b>METHODS</b>Human umbilical venous endothelial cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with different treatments including the normal glucose (5.5 mmol/L), high glucose (33 mmol/L) and aspirin (0.01 - 1.00 mmol/L) with high glucose. And 300 micromol/L L-NAME was added to the culture medium when needed. After 48 hours, SA-beta-gal staining was used to evaluate the senescence. Total nitric oxide (NO) production and NO synthase (NOS) activity were measured using Griess reaction and molecular probes of 3-amino-4-aminomethyl-2', 7'-difluorescein, diacetate. The level of intracellular reactive oxygen species was monitored by flow cytometry using 2', 7'-dichlorofluorescein diacetate. Endothelial NOS (eNOS), caveolin-1 protein expressions and caveolin-1/eNOS interaction were analyzed by immunoblotting and immunoprecipitation respectively. Asymmetric dimethylarginine (ADMA) concentration was determined by high-performance liquid chromatography.</p><p><b>RESULTS</b>Exposure to 33 mmol/L glucose for 48 hours significantly increased the number of SA-beta-gal positive cells. Co-incubation with aspirin markedly inhibited SA-beta-gal activity dose-dependently. Aspirin increased NOS activity with eNOS protein expression unchanged and increased NO levels and alleviated oxidative stress. Consistent with these findings, caveolin-1 expression, caveolin-1/eNOS interaction and ADMA accumulation were also decreased. All the inhibitory effects of aspirin on senescence were completely obliterated by L-NAME, the NOS inhibitor.</p><p><b>CONCLUSION</b>The anti-senescent effects of aspirin are fulfilled by increasing NO production via the up-regulation of NOS activity and preventing caveolin-1 expression, caveolin-1/eNOS interaction and ADMA accumulation.</p>

N-ras and fms gene mutation in idiopathic thrombocytopenic purpura and myelodysplasia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the pathogenesis of idiopathic thrombocytopenic purpura (ITP) and improve the differential diagnosis from myelodysplastic syndromes (MDS).</p><p><b>METHODS</b>Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was performed to detect the point mutation of codon 12,13 in N-ras gene and codon 301, 969 in fms gene in adult and aged ITP and MDS patients.</p><p><b>RESULTS</b>In 25 ITP patients, N-ras mutation and fms mutation were detected in one each (4%). Mutations were found in 3 of 8 MDS patients: two (25%) with N-ras mutation and one (12.5%) with fms mutation.</p><p><b>CONCLUSIONS</b>Patients with N-ras or fms gene mutation diagnosed as MDS rather than ITP.</p>

Preoperative intra-arterial chemotherapy for progressive lower rectal cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of preoperative regional intra-arterial chemotherapy (PRAC) on progressive lower rectal cancer.</p><p><b>METHODS</b>Forty-five patients with progressive lower rectal cancer were divided into groups A (23 cases) and B (22 cases) for treatment with PRAC 1 to 2 weeks prior to surgical tumor resection or with surgical resection only, respectively.</p><p><b>RESULTS</b>PRAC caused obvious tissue degeneration and necrosis of rectal cancer with a total effective rate of 95.65%. The rates of radical resection in groups A and B were 91.3% and 72.27%, respectively. The 1-year postoperative survival rates of the two groups were 95.65% and 86.36%, with 3-year survival of 89.96% and 68.18%, and 3-year postoperative recurrence rates of 8.69% and 27.27%, respectively. The anal preservation rates of the two groups were 78.26% and 59.09%.</p><p><b>CONCLUSION</b>PRAC can increase radical resection rates, promote the postoperative survival and anal preservation rate, and lower the recurrence rate in patients with lower rectal cancer.</p>

Expression and purification of human amelogenin in Escherichia coli / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To establish the expression and purification route for the gene encoding human amelongenin (AMG) mature peptide in Escherichia coli (E. coli).</p><p><b>METHODS</b>Recombined plasmid pGEX-4T-1/AMG was identified by double endonuclease digestion electrophoretogram and DNA sequence analysis. The recombined plasmid was transformed to E. coli BL21. The inducing time, isopropyl-beta-D-thiogalactopyranoside (IPTG) concentration and inducing temperature were optimized for the express system. Under the optimized condition, the target fusing protein in superatant, periplasm, plasm and inclusion body was analyzed separately. A great amount of target fusing protein was found in the dissoluble protein. AMG fusing protein was purified by the GSTrapFF affinity column.</p><p><b>RESULTS</b>Double endonuclease digestion electrophoretogram and DNA sequence analysis were done to identify the recombined vector pGEX-4T-1/AMG. The results were consistent with the anticipation. The optimum inducing time was 14.5 hours. The optimum IPTG concentration was 1.0 mmol/L. The optimum inducing temperature was 20 degrees C. Under this condition, the target protein was expressed to a maximum. Plentiful target protein was expressed in plasm and inclusion body under the optimized condition. A mount of plasm protein was obtained and purified by the GSTrapFF affinity column. The purified liquid was collected and analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PACE). The protein electrophoresis map showed that AMG fusing protein was purified successfully. After twice elution, high pure fusing protein was obtained.</p><p><b>CONCLUSION</b>pGEX-4T-1/AMG system is used successfully to express human AMG fusing protein.</p>