Objectives To investigate the expression of 8-Hydroxy-2'-deoxyguanine(8-oxodG)in white blood cell,plasma and urine of rhesus monkey of different age group.Methods 30 female rhesus macaques at different age(1y,5y,10y,15y,20y,25y)were selected and grouped(n=5,each).10 mL of morning urine and 5 mL of fasting venous blood were collected.The level of 8-oxodG expression in plasma,leukocyte and urine was measured by high-performance liquid chromatography-mass spectrometry(HPLC-MS) method.Results The level of 8-oxodG in leukocytes,plasma and urine was increased along with aging.The level of 8-oxodG was 1.8,1.6 and 1.4 times higher in 25 year group than in 1 year group in plasma,white blood cell and urine,respectively(P＜0.05).The 8-oxodG level was more than 40 times higher in urine than in plasma.Conclusions The expression level of 8-oxodG is increased along with aging.It may be one of the experimental evidence of the aging markers.

BACKGROUND:Stem celltumorigenicity is a practical issue concerning stability in the clinical application of stem cells. Therefore, it is particularly important to clear whether stem cells have tumorigenic ability or not. Nude mice occupy an increasingly important position in oncology, immunology, and safety evaluation of drugs and biological products.
OBJECTIVE:To observe the tumorigenicity of neural stem cells and mesenchymal stem cells in Balb/c nude immunodeficient mice.
METHODS:Balb/c nude mice were randomly divided into control group, negative group, positive group, neural stem cellgroup and mesenchymal stem cellgroup. HepG-2 cells, RPE cells, passage 4 neural stem cells and mesenchymal stem cells were injected subcutaneously into nude mice from different groups, respectively. After 12 weeks of injection, anatomical observation was performed to detect the tumor formation in the transplantation site. Meanwhile, soft agar colony formation assay was applied to investigate neural stem celland mesenchymal stem cellclone in vitro.
RESULTS AND CONCLUSION:After 12 weeks of injection, the tumorigenicity study results showed that no tumor developed in the transplantation site in the control group, negative group, neural stem cellgroup and mesenchymal stem cellgroup. Histopathologic examinations also showed no abnormality in these groups. Soft agar colony formation assay showed in soft agar resistance medium, neural stem cells and mesenchymal stem cells did not exhibit clone ability. These findings indicate that neural stem cells and mesenchymal stem cells undergoing short-term passages have no tumorigenic growth.

BACKGROUND:Stem cells expansion technology in vitro is affected by the microenvironment of the culture system. So, to find an effective method is particularly important to promote celladherent and growth. OBJECTIVE:To compare the effects of different culture media on cellexpansion. METHODS:Human umbilical cord mesenchymal stem cells at a density of 5.0×104 cells/cm2 were inoculated into Corning? polystyrene culture dishes coated with or without poly-L-ornithine and Corning? cellBIND culture dishes. celladhesion and proliferation were observed, and expressions of celladhesion proteins and cellmarkers were detected. RESULTS AND CONCLUSION:celladhesion was promoted when cells were cultured in Corning? cellBIND? Surface medium coated with poly-L-ornithine for 24 hours, and the cultured cells grew at the logarithmic phase. The cellproliferation was also enhanced, and the cells expressed celladhesion protein but not the cellmarkers of CD73, CD90, CD105. Corning? cellBIND? Surface medium was superior to Corning? polystyrene culture medium in the improvement of celladhesion and proliferation. Additional y, both of these two media showed no influence on cellphenotype. These findings indicate that Corning? cellBIND? Surface medium can promote celladhesion and proliferation, but shows no effects on cyclin and cellphenotype. This medium coated with poly-L-ornithine can further accelerate celladhesion and proliferation, and stably express cellphenotype of human umbilical cord mesenchymal stem cells.

Objective To study the changes in protein expression levels of 8 oxide of guanine in the temporal and frontal lobes in SAMP8 (senescence accelerated mice)mice versus SAMR1 (control mice).Methods In the study,we collected 12 SAMP8 mice and 12 SAMR1 mice at different month stage(1m,4m,8m,12m)(n=6,each group).After anesthesia,mice were sacrificed.The temporal and frontal lobe tissues of mice were labeled according to the stereotactic anatomical map of mouse brain and taken up for study.After paraffin sections were made,the expression level of 8 oxide of guanine in brain tissues was detected by immunohistochemical method.Results In the same age group at 1 month in the same temporal lobe,the average optical density of 8-oxodG was slightly higher in SAMP8 mice(0.086)than in SAMR1 mice(0.061,t =1.541,P＞0.05).There were significant differences in the mean optical density of 8-oxodG in temporal lobe in 4,8 and 12 months groups between SAMP8 and SAMR1 group(0.101 vs.0.081,0.147 vs.0.109 and 0.176 vs.0.120,t =2.405,2.612 and 5.019,respectively,P ＜0.05).There was no significant difference in the average optical density levels of 8-oxodG in frontal lobe in 1 and 4 months groups between SAMP8 and SAMR1 mice (0.044 vs.0.030,0.062 vs.0.046,t =1.843 and 1.163,respectively,P ＞ 0.05),while the average optical density levels of 8-oxodG in 8 and 12 months groups(0.090 and 0.138)were higher than those of SAMR1 mice in the same age group(0.049 and 0.063)(t =5.327 and 4.88,P＜0.01).There was no significant difference in the average optical density of 8-oxoG in temporal lobe in 1 month group between SAMP8 and SAMR1 mice(0.099 vs.0.066,t =1.956,P ＞0.05).There were significant differences in the average optical density levels of 8-oxoG in temporal lobe in 4,8 and 12 months groups between SAMP8 and SAMR1 mice(0.119 vs.0.076,0.148 vs.0.094 and 0.173 vs.0.101,t =4.033,3.042 and 5.328,respectively,P＜0.01).There was no significant difference in the average optical density levels of 8-oxoG in frontal lobe in 1 month and 4 months groups between SAMP8 and SAMR1 mice(0.049 vs.0.039,0.058 vs.0.056,t =0.713 and 0.172,respectively,P ＞0.05).The average optical density of 8-oxoG in frontal lobe in 8 months and 12 months groups had significant differences between SAMP8 and SAMR1 mice(0.087 vs.0.058,0.12 vs.0.063,t =2.261 and 4.185,P ＜0.05).In addition,we also found that the average optical density of 8 oxide of guanine in same SAMP8 mice at the same age was higher in temporal lobe than in frontal lobe.Conclusions The protein levels of 8-oxodG and 8-oxoG are increased with age,and the damage caused by nucleic acid oxidation is more severe in temporal lobe than in frontal lobe in SAMP8 mice.

This study was purposed to investigate the effects of N-Arachidonoylethanolamine (ANA) on the quality of platelets (Plt) stored in Plt M-sol preservative solution at 22 ± 2°C. Samples taken from collecting apheresis Plt by the Amicus instrument and splited into two equal parts were stored in Plt M-sol preservative solution on a shaker at 22 ± 2°C. Different working concentrations of ANA (from 0.1 to 50 µmol/L) were then added into one part of stored Plt as the experimental group, the other without ANA was used as the control group. The viability of Plts stored at 22 ± 2°C for 7 days was evaluated by MTT colorimetric assay. The most effective concentration of ANA was selected and added to the subsequent experimental group. Plt count (BPC), mean Plt volume (MPV), Plt distribution width (PDW), phosphatidyl serine (PS) and soluble P-selectin were detected on the 1(st), 5(th), 7(th), 9(th) and 11(th) day of storage. The results showed that the most effective working concentration of ANA was 0.5 µmol/L, which showed significant increasing Plt viability (91.23 ± 5.44%) compared to the control group (62.54 ± 4.79%). Thus, ANA concentration at 0.5 µmol/L was choose to perform subsequent experiments. During 11 days of storage, the BPC, MPV and PDW were not changed significantly between the experimental group and control group, although there was decreasing trend in the BPC and increasing trends in MPV and PDW in the two groups. The rate of Plt PS positive was enhanced during the storage period: the rate of PS positive in experimental group increased from 7.69 ± 1.82% to 10.74 ± 1.78% while it in control group increased from 11.21 ± 2.03% to 15.37 ± 1.95%, with significant differences between the two groups (P < 0.05) on the 9(th) and 11(th) day of storage, respectively. Soluble P-selectin contents in experimental group on the 9(th) and 11(th) day of storage were 30.19 ± 2.03 ng/ml and 34.52 ± 2.64 ng/mL, respectively, while those in control group were 39.18 ± 2.66 ng/ml and 43.23 ± 2.58 ng/ml, respectively, with significant differences between the two groups (P < 0.05). It is concluded that the extended storage of Plt in M-sol treated with low concentration ANA can potentially alleviate Plt storage lesions.
Adult ; Blood Platelets ; drug effects ; Blood Preservation ; Endocannabinoids ; pharmacology ; Female ; Humans ; Male

This study was aimed to identify a novel HLA-DRB1 allele from a Chinese potential hemopoietic stem cell donor of Northeast China. A rare HLA-DRB1 allele was initially detected by Luminex PCR-SSO typing, then the sample was sequenced by sequence-based typing (SBT) and the alignments of sample's alleles was identified by single allele-specific sequencing strategy. The results revealed the existence of a new allele which differs from the closest matching allele DRB1*03:06 by a single nucleotide substitution at position 239, where C→G in exon 2, resulting in an amino acid exchange from Thr to Arg at codon 51. It is concluded that a novel allele has been confirmed and its name DRB1*03:80 is officially assigned by the WHO Nomenclature Committee in February 2012.
Alleles ; Asian Continental Ancestry Group ; genetics ; HLA-DRB1 Chains ; genetics ; Humans ; Male ; Sequence Analysis, DNA

OBJECTIVE: To study the Polymorphism of the human platelet antigen(HPA) gene 1-17 and human leukocyte antigen(HLA) gene-A and B locus in Shandong Han population. METHODS: A total of 962 samples from routine voluntary platelet donors were genotyped for HPA1-17 system and HLA-A site, B by PCR-SSP and PCR-SSOP respectively．Gene frequencies were calculated by counting. HPA1-17 and HLA genotype combinations were analyzed by Arelequin 3.5. RESULTS: The gene frequencies of HPA-la, -1b, HPA-2a, -2b, HPA-3a, -3b, HPA-4a, -4b, HPA-5a, -5b, HPA-6a, -6b, HPA-15a, -15b were 0.9918, 0.0082, 0.9419, 0.0592, 0.5841, 0.4174, 0.9969, 0.0031, 0.9892, 0.0108, 0.9835, 0.0175,0.5488 and 0.4512, respectively． The most common HPA genotype combination was HPA-(1, 2, 4, 5, 6, 7-14, 16, 17) aa-3ab-15ab (0.2048). Moreover, HLA-A*2(0.3094) and HLA-B*13(0.1513) showed the highest frequency in their respective locus. The most common HLA genotype combination was HLA-A*2-B*13(0.1397) . CONCLUSION Distributions of HPA and HLA show high polymorphism in Shandong Han population. The ethnic and territorial difference of HPA distribution is also confirmed. It is imperative to establish local genetic database of volunteer platelet donors.
Alleles ; Antigens, Human Platelet/genetics* ; Gene Frequency ; Genotype ; Humans ; Polymorphism, Genetic

Objective: To investigate hepatic perivascular epithelioid cell tumor (PEComa) diagnosis and treatment plan. Methods: 24 cases diagnosed with PEComa clinical manifestations, serum alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA199), imaging findings, surgical methods, postoperative hospital stay, pathological results and prognosis were analyzed retrospectively from September 2015 to September 2020. Results: Majority of patients were females (79.2%), aged 41.5±11.4 years. Tumors were predominantly located in the right liver (50.0%). 76.7% of the cases were mostly clinically asymptomatic. AFP, CEA and CA199 indices were all negative. CT mostly showed low density tumor in the plain scan phase, enhanced in the enhancement phase, and enhanced and weakened in portal venous and equilibrium phase (66.7%). MRI manifestations of most tumors were hypointense on T₁WI and hyperintense on T₂WI (72.7%). B-ultrasound mostly showed hyperechoic mass in the tumor area with punctate vascular shadow (52.9%). Postoperative hospital stay was 9.0±2.4 days for laparoscopic surgery patients (n=13), 13.4±6.3 days for traditional laparotomy (hereinafter referred to as laparotomy, n=10), and 3 days for 1 patient with microwave ablation. All postoperative pathological results were positive for HMB45 and Melan-A. Follow-up results: 21 cases survived normally, with no tumor recurrence in the recent physical examination; two cases had tumor recurrence and they died two and three years after surgery, and one case was lost to follow-up. Conclusion: Hepatic PEComa more commonly occurs in middle-aged women, with no specific features for tumor markers and clinical manifestations. Some imaging findings are specific, so its features can be combined as a basis for diagnosis. Postoperative pathological examination results can confirm the diagnosis. Therefore, surgery remains the initial treatment plan. Microwave ablation and laparoscopic surgery are recommended as the preferred option because of shorter hospital stays and less trauma than open surgery.
Adult ; Biomarkers, Tumor/analysis* ; Carbohydrates ; Carcinoembryonic Antigen ; Female ; Humans ; Liver/pathology* ; MART-1 Antigen ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Perivascular Epithelioid Cell Neoplasms/surgery* ; Retrospective Studies ; alpha-Fetoproteins

ObjectiveTo investigate the drug-resistant gene polymorphisms in Plasmodium falciparum imported from Equatorial Guinea to Shandong Province. MethodsFrom 2015 to 2016, blood samples were collected from imported P. falciparum malaria patients returning from Equatorial Guinea to Shandong Province, and genome DNA of the malaria parasite was extracted. The drug-resistant Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum were amplified using a PCR assay, followed by DNA sequencing, and the sequences were aligned. Results The target fragments of all 5 drug-resistant genes of P. falciparum were successfully amplified and sequenced. There were 72.8%, 18.6%, and 8.6% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfcrt gene, respectively, and all mutant haplotypes were CVIET (the underline indicates the mutation site). There were 20.0%, 61.4% and 18.6% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfmdr1 gene, respectively, and the mutant haplotypes mainly included YF and NF (the underlines indicate the mutation sites). There were 1.4%, 98.6%, and 0 of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfdhfr gene, respectively, and AIRNI was the predominant mutant haplotype (the underline indicates the mutation site). There were 1.4%, 94.3%, and 4.3% of P. falciparum parasites carrying the wild-, mutant-, and mixed-type Pfdhps gene, respectively, and SGKAA was the predominant mutant haplotype (the underline indicates the mutation site). The complete drug-resistant IRNGE genotype consisted of 8.6% of the Pfdhfr and Pfdhps genes, and the K13 gene A578S mutation occurred in 1.4% of the parasite samples. Conclusions There are mutations in the Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, and K13 genes of P. falciparum imported from Equatorial Guinea to Shandong Province, with a low frequency in the Pfcrt gene mutation and a high frequency in the Pfmdr1, Pfdhfr, and Pfdhps gene mutations, and the K13 gene A578S mutation is detected in the parasite samples.

OBJECTIVE: To examine whether the combination of Naoxintong Capsule with standard care could further reduce the recurrence of ischemic stroke without increasing the risk of severe bleeding. METHODS: A total of 23 Chinese medical centers participated in this trial. Adult patients with a history of ischemic stroke were randomly assigned in a 1:1 ratio using a block design to receive either Naoxintong Capsule (1.2 g orally, twice a day) or placebo in addition to standard care. The primary endpoint was recurrence of ischemic stroke within 2 years. Secondary outcomes included myocardial infarction, death due to recurrent ischemic stroke, and all-cause mortality. The safety of drugs was monitored. Results were analyzed using the intention-to-treat principle. RESULTS: A total of 2,200 patients were enrolled from March 2015 to March 2016, of whom 143 and 158 in the Naoxintong and placebo groups were lost to follow-up, respectively. Compared with the placebo group, the recurrence rate of ischemic stroke within 2 years was significantly lower in the Naoxintong group [6.5% vs. 9.5%, hazard ratio (HR): 0.665, 95% confidence interval (CI): 0.492-0.899, P=0.008]. The two groups showed no significant differences in the secondary outcomes and safety, including rates of severe hemorrhage, cerebral hemorrhage and subarachnoid hemorrhage (P>0.05). CONCLUSION The combination of Naoxintong Capsule with standard care reduced the 2-year stroke recurrence rate in patients with ischemic stroke without increasing the risk of severe hemorrhage in high-risk patients. (Trial registration No. NCT02334969).
Adult ; Humans ; Secondary Prevention/methods* ; Ischemic Stroke ; Stroke/prevention & control* ; Cerebral Hemorrhage/complications* ; Double-Blind Method ; Platelet Aggregation Inhibitors