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OBJECTIVE@#To observe the clinical effect of high suspension and low incision (HSLI) surgery on mixed haemorrhoids, compared with Milligan-Morgan haemorrhoidectomy.@*METHODS@#A multi-centre, randomized, single-blind, non-inferiority clinical trial was performed. Participants with mixed haemorrhoids from Xiyuan Hospital of China Academy of Chinese Medical Sciences, Beijing Rectum Hospital, Air Force Medical Center of People's Liberation Army of China, and Puyang Hospital of Traditional Chinese Medicine were enrolled from September 2016 to March 2018. By using a blocked randomization scheme, participants were assigned to two groups. The experimental group was treated with HSLI, while the control group was treated with Milligan-Morgan haemorrhoidectomy. The primary outcome was the clinical effect evaluated at 12 weeks after operation. The secondary outcomes included the number of haemorrhoids treated during the operation, pain scores, use of analgesics, postoperative oedema, wound healing, incidence of anal stenosis, anorectal manometry after operation, as well as surgical duration, length of stay and total hospitalization expenses. A safety evaluation was also conducted.@*RESULTS@#In total, 246 eligible participants were enrolled, with 123 cases in each group. There was no significant difference in the clinical effect between the two groups (100.00% vs. 99.19%, P>0.05). Compared with the control group, the number of external haemorrhoids treated during the operation and the pain scores after operation were significantly reduced in the experimental group (P0.05). The surgical duration and length of stay in the experimental group were significantly longer than those in the control group, and the total hospitalization expense was significantly higher than that in the control group (all P<0.05). No adverse events were reported in either group during the whole trial or follow-up period.@*CONCLUSION@#HSLI had the advantages of preserving the skin of anal canal completely, alleviating postsurgical pain and promoting rapid recovery after operation. (Registration No. ChiCTR1900022883).
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OBJECTIVE: To investigate the effects of nuclear transcription factor-κB(NF-κB)/amide adenine dinucleotide phosphate oxidase 1(NOX1) signaling pathway in tumor necrosis factor-α(TNF-α) induced apoptosis of A549 cells. METHODS: i) A549 cells were stimulated with TNF-α at the concentrations of 0.00, 0.25, 0.50, and 1.00 nmol/L. CCK-8 assay was used to detect the cell viability to screen the optimal stimulating concentration of TNF-α. ii) A549 cells at logarithmic growth stage were randomly divided into four groups, the control group, the TNF-α group, the BAY11-7082(NF-κB inhibitor) group and the TNF-α+BAY11-7082 group. The cells in the control group were not treated. The TNF-α and BAY11-7082 groups were stimulated with 0.50 nmol/L TNF-α and 5 μmol/L BAY11-7082, respectively. The TNF-α+BAY11-7082 group was stimulated by both TNF-α and BAY11-7082. After 24 hours of culture, the cell survival rate was detected by CCK-8 assay. Flow cytometry was used to detect cell apoptotic rate, and Western blotting was used to detect the relative expression of NF-κB(p65) and NOX1 proteins. RESULTS: i) When A549 cells were stimulated with TNF-α at the concentration of 0.50 nmol/L, the cell proliferative activity was reduced and the cell apoptosis was promoted. This concentration was selected as the stimulation dose of TNF-α in subsequent experiments. ii) The survival rate of A549 cells in the TNF-α group decreased(P<0.05), the apoptotic rate and the protein expressions of NF-κB(p65) and NOX1 increased in TNF-α group(all P<0.05) compared with the control group. In BAY11-7082 group, the survival rate and the relative expression of NF-κB(p65) and NOX1 of A549 cells were decreased(all P<0.05), and the apoptotic rate of A549 cells was increased(P<0.05) compared with the control group. A549 cells in TNF-α+BAY11-7082 group changed from a long spindle shape to an irregular one. The cell survival rate increased(P<0.05), the apoptotic rate and the relative expression of NF-κB(p65) and NOX1 decreased(all P<0.05) compared with the TNF-α group. CONCLUSION: NF-κB/NOX1 signaling pathway is involved in A549 cells apoptosis induced by TNF-α.
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Objective To evaluate the AChE and nAChR in the NMJ and the morphology of the muscle in the bilateral triceps surae after the unilateral shock wave therapy. Method 60 male New Zealand rabbits weighing (2± 0.2) Kg were used in this study. Two thousand shock waves at an energy flux density of 1.5bar and the frequency of 10Hz were applied to their left calf muscles. Divided into six groups, both sides of the triceps muscle of calf were taken out on the day of the shock wave and the1,2,4,6 and 8 weeks after the treatment. HE staining was used to observe the morphological changes of muscle tissue and the average optical density was measured after AChE stain so as to calculate the receptor count after Acetylcholine receptor immunohistochemistry. Result No abnormal morphological abnormalities were observed in all rabbits. In the first five groups, the AChE was significantly higher in the side of the shockwave treatment compairing with the control side (<0.05),slow decrease after 1 week after the treatment. In the first five groups, the nAChR was significantly lower in the side of the shock wave treatment compairing with the control side ( <0.05), and gradually increased to normal after 8 weeks. Conslusion Suitable dose of shock wave will not have a greater impact on morphology of muscle tissue. After the shock wave treatment, the amount and degree of stimulate of muscle cells were decreased, and the production of action potentials was reduced. While the experimental side AchE and AchR in shock wave treatment day to 8 weeks after treatment showed a significant trend to normal, it shows that the effect of shock wave on NMJ is transient and reversible.
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Purpose To summarize the histological,immunophenotypic feature and MYCN (MYC gene) amplification results of peripheral neuroblastic tumours in children,and to predict its biological behavior and prognosis.Methods The histology and immunophenotype of 100 cases of neuroblastoma (NB) and ganglioneuroblastoma (GNB) were retrospectively analysed,MYCN status was detected in 60 cases.Results Among The average age of 100 cases of GN and GNB was 2.7 years old,and that of males was more than that of females.NB could be divided into three subtypes:undifferentiated,poorly differentiated and differentiating.GNB could be divided into two subtypes:intermixed (iGNB) and nodular (nGNB).Immunohistochemical staining showed neuroblastoma cells were positive for NSE,NF,PGP9.5,Syn,CgA in varying degree.Schwann cells were positive for S-100 and GFAP.MYCN amplification was detected in 1 1.67％ of the cases,and no MYCN amplification was seen in iGNB patients.Conclusion The diagnosis of peripheral neuroblastic tumours is mainly based on histological morphology,special tests (immunohistochemistry,electron microscope and cytogenetics) can helpful for identifing undifferentiated neuroblastoma cells.The prognosis of neuroblastoma derived tumors is evaluated according to the age of patients,the classification and staging of tumors,and molecular genetic alterations.
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Objective To examine the effect of eating behavior and dietary intake on the sleep quality of college students,and to provide basis for improving students' sleep quality.Methods College students from Hangzhou Normal University were randomly selected and they were investigated from October to November,2014.Pittsburgh Sleep Quality Index (PSQI) was used to measure the quality of sleep,and a self-designed questionnaire was used to measure eating behavior and dietary intake.Results A total of 588 college students were investigated.The average score of PSQI was 4.72 ±2.52,and 76 (12.9％) of the respondents have poor sleep quality.Single-factor analyses showed that sleep quality was not associated with gender,BMI,cigarette smoking and academic pressure,but was associated with peer effect,meal regularity,and frequency of fish and poultry intake (P ＜ 0.05).Multi-factor logistic regression showed that frequency of eating-out (OR =3.04,95％ CI:1.58-5.84;P =0.001,4 or more than 4 times vs.less than once per week) irregular dinner (OR =1.96,95％ CI:1.23-3.40;P =0.017,irregular vs.regular) and less fish intake (OR =2.48,95％ CI:1.27-4.85;P =0.01,less than once vs.2-3 times per week) increased the risk of poor sleep quality.Conclusion Eating behavior and dietary intake are closely related to sleep quality of college students and they should concern about meal regularity and nutrition balance.
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<p><b>OBJECTIVE</b>To explore the biological mechanisms underlying Angelica sindsis polysaccharide (ASP) -induced aging of human-derived leukemia stem cells (LSCs) in vitro.</p><p><b>METHOD</b>Acute myelogenous leukemia stem cells were isolated by magnetic activated cell sorting (MACS). The ability of LSC proliferation treated by various concentration of ASP(20-80 mg · L(-1)) in vitro for 48 hours were tested using cell counting Kit-8 ( CCK8) , colony forming were evaluated by methylcellulose CFU assay. The ultra structure changes of AML CD34+ CD38- cells were analyzed by transmission electron microscopy. The aging cells were detected with senescence-β-galactosidase Kit staining. Expression of aging-related p53, p21, p16, Rb mRNA and P16, Rb, CDK4 and Cyclin E protein were detected by quantitative reverse transcription polymerase chain reaction( qRT-PCR) and Western blotting, respectively.</p><p><b>RESULT</b>The purity of the CD34 + CD38 - cells is (91.15 ± 2.41)% after sorted and showed good morphology. The proliferation of LSC was exhibited significantly concentration-dependent inhibited after exposure to various concentration of ASP. Treated by 40 mg · L(-1) ASP for 48 hours, the percentage of positive cells stained by SA-β-Gal was dramatically increased (P < 0.01) and the colony-formed ability has been weakened (P < 0.01). The observation of ultrastructure showed that cell heterochromatin condensation and fragmentation, mitochondrial swelling, lysosomes increased in number. Aging-related p53, p21, p16, Rb and P16, Rb were up-regulated, protein regulatory cell-cycle CDK4 and Cyclin E were down-regulated. ASP may induce the senescence of LSCs effectively in vitro, P16-Rb cell signaling pathway play a significant role in this process.</p><p><b>CONCLUSION</b>ASP can induce human leukemia stem cell senescence in vitro, the mechanism involved may be related to ASP regulation P16-Rb signaling pathways.</p>
Subject(s)
Humans , Angelica sinensis , Chemistry , Cell Cycle , Cell Cycle Proteins , Genetics , Metabolism , Cells, Cultured , Cellular Senescence , Drugs, Chinese Herbal , Pharmacology , Gene Expression Regulation, Leukemic , Leukemia , Drug Therapy , Genetics , Metabolism , Neoplastic Stem Cells , Cell Biology , Polysaccharides , Pharmacology , Signal TransductionABSTRACT
Objective: To explore the effects and the possible mechanism of combined injection of Angelica sinensis polysaccharide (ASP) and cytarabine (Ara-C) on the bone marrow mononuclear cells (BMMCs) of the transplanted human leukemia mouse model. Methods: K562 cells (2×107)were transplanted into the tail vein of mice to establish the transplanted human leukemia NOD/SCID mouse model. Then the leukemia mice were randomly divided into model, ASP, Ara-C, and ASP + Ara-C groups. After the 30 d transplantation, the mice were ip injected with ASP (200 mg/kg/d), Ara-C (2.5 mg/kg/d), and ASP (200 mg/kg/d) + Ara-C (2.5 mg/kg/d), respectively for 14 d, and the mice in the model group were injected with saline (equal volume and time). The next day after the treatment, the eyeball blood was collected to detect the amount and classification of white blood cells (WBC). The femurs were taken to count BMMCs of each femur. The proliferation of BMMCs was detected by CCK-8; The distribution of cell cycles was analyzed by flow cytometry (FCM); The capability of colony forming was examined by CFU-Mix cultivation; The ratio of the SA-β-Gal staining positive BMMCs was counted; The aging related proteins of P16, Rb, CDK4, and CyclinD1 were detected by Western blotting. Results: Compared with the model group, ASP or Ara-C injected alone and their combined injection can obviously reduce the amount of the peripheral blood WBC, the percentage of neutrophiles, and the number of femur BMMCs; effectively inhibit the proliferation of BMMCs, CFU-Mix forming, and the ratio of S stages; markedly raise the percentage of lymphocytes, ratio of G1 stages, and the percentage of SA-β-Gal positive cells; down-regulate the expression of the aging related proteins of CDK4 and CyclinD1; and up-regulate the expression of P16 and Rb protein. The effects of ASP + Ara-C group were much better than those in the other groups. Conclusion: The aging mechanism of BMMCs for the transplanted human leukemia mice induced by ASP and Ara-C may be ascribed to the regulation of the expression of the aging related proteins of P16, Rb, CDK4, and CyclinD1. Our research provides a new idea to treat leukemia in clinic.
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The latest findings of our laboratory showed that Angelica sinensis polysaccharide (ASP) showed a definite effect in regulating the aging of hematopoietic stem cells. Leukemia is a type of malignant hematopoietic tumor in hematopoietic stem cells. There have been no relevant reports about ASP's effect in regulating the aging of leukemia cells. In this study, human acute myeloid leukemia (AML) KG1alpha cell lines in logarithmic growth phase were taken as the study object, and were divided into the ASP group, the cytarabine (Ara-C) group, the ASP + Ara-C group and the control group. The groups were respectively treated with different concentration of ASP, Ara-C and ASP + Ara-C for different periods, with the aim to study the effect of ASP combined with Ara-C in regulating the aging of human acute myeloid leukemia KG1alpha cell lines and its relevant mechanism. The results showed that ASP, Ara-C and ASP + Ara-C could obviously inhibit KG1alpha cell proliferation in vitro, block the cells in G0/G1 phase. The cells showed the aging morphological feature. The percentage of positive stained aging cells was dramatically increased, and could significantly up-regulate the expression of aging-related proteins P16 and RB, which were more obvious in the ASP + Ara-C group. In conclusion, the aging mechanism of KG1alpha cell induced by ASP and Ara-C may be related to the regulation of the expression of aging-related proteins, suggesting that the combined administration of ASP and anticancer drugs plays a better role in the treatment of leukemia .
Subject(s)
Humans , Aging , Genetics , Metabolism , Angelica sinensis , Chemistry , Cell Cycle , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Metabolism , Leukemia , Drug Therapy , Genetics , Metabolism , Polysaccharides , Pharmacology , Retinoblastoma Protein , Genetics , Metabolism , Tumor Cells, CulturedABSTRACT
Leukemia is a type of malignant tumors of hematopoietic system with the abnormal increased immature leukemia cells showing metastasis and invasion ability. Liver is one of the main targets of the leukemia cells spread to, where they may continue to proliferate and differentiate and cause liver function damage, even liver failure. Our previous studies showed that Angelica polysscharides (APS), the main effective components in Angelica sinensis of Chinese traditional medicine, was able to inhibit the proliferation and induced differentiation of the leukemia cells, however, its effect on the liver during the treatment remains elucidated. In the present study, the human leukemia NOD/SCID mouse model were established by implantation human leukemia K562 cells line, then the leukemia mouse were treated with APS, Ara-c or APS + Ara-c respectively by peritoneal injection for 14 days, to explore the effect and mechanism of the chemicals on the mouse liver. Compared to the human leukemia NOD/SCID mouse model group with the treatments of APS, Ara-c and APS + Ara-c, We found that severe liver damage and pathological changes of the liver were able to alleviate: First, the number of white blood cells in the peripheral blood was significantly lower and with less transplanted K562 leukemia cells; Second, liver function damage was alleviated as liver function tests showed that alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were significantly reduced, while the albumin (Alb) was notably increased; Third, liver antioxidant ability was improved as the activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly increased, and the contents of GSH and malonaldehyde (MDA) were decreased significantly in the liver; Fourth, the inflammation of the liver was relieved as the level of IL-1beta and IL-6, the inflammatory cytokines, were decreased significantly in the liver. Fifth, liver index was increased as the pathological observation showed that leukemia cells with diffused infiltration into the liver lobules were significantly reduced and with a remarkable increase of apoptotic positive cell rate by TUNEL test. Furthermore, the APS + Ara-c combined administration showed an even more significant positive effect. In conclusion, the APS, Ara-c therapy reduced the accumulation of leukemia cells within the liver, reduced the liver function damage and levels of inflammatory factors, improved antioxidant capacity of the liver tissue and thus alleviate the pathological changes of the liver. Moreover, the APS + Ara-c combination therapy may have an additive effect.
Subject(s)
Animals , Humans , Male , Mice , Angelica , Chemistry , Antineoplastic Combined Chemotherapy Protocols , Pharmacology , Cell Line, Tumor , Cytarabine , K562 Cells , Leukemia , Drug Therapy , Liver , Mice, SCID , PolysaccharidesABSTRACT
<p><b>OBJECTIVE</b>The effect of angelica sinensis polysaccharides (ASP) on the production of reactive oxygen specie (ROS), the capability of total anti-oxidant (T-AOC), and the expression of p16 in mRNA level in mice hematopoietic stem cells (HSCs) were observed to explore the underlying mechanism that ASP delay aging of HSCs in vivo.</p><p><b>METHOD</b>C57BL/6J mice were randomly divided into normal group, aging group, and the above groups treated with ASP. Mice were uniformly explored in X-ray (3.0 Gy/8 F) to erect model of aging. Normal and aging ASP intervention groups mice were treated with ASP by intragastric administration, while normal and aging groups were treated with equal-volume NS during X-ray irradiation. Mice HSCs were isolated by magnetic cell sorting and cultured in vitro. Senescence-associated beta-galactosidase (SA-beta-Gal) staining was used to detect aging HSCs. Cell cycles analysis and CFU-Mix cultivation were used to evaluate the capability of self-renewing and colony forming in HSCs. The production of ROS in HSCs was evaluated by flow cytometry analysis and immunofluorescence assess, respectively. T-AOC was detected by chemical colorimetric method. The expression of p16 was determined by real-time quantitative PCR (qRT-PCR).</p><p><b>RESULT</b>Exogenous X-ray irradiation induced HSCs aging was compared with normal group without irradiation. Biological feature of HSCs in aging group with X-ray irradiation as follows: The percentage of SA-beta-Gal positive cells, the ratio of G1 stages and the production of ROS were significantly increased , the expression of p16 in mRNA level was also upregulated. The capacility of colony forming and T-AOC in HSCs were decreased. ASP could significantly decrease the percentage of SA-beta-Gal positive cells, the ratio of G1 stages and the production of ROS in HSCs, and downregulate the expression of p16 in mRNA level in HSCs contrast to aging group without ASP treatment. In addition, ASP could remarkably increase T-AOC and the capacility of colony forming in HSCs compared with aging group without ASP treatment.</p><p><b>CONCLUSION</b>X-ray (3.0 Gy/8 F) could induce mice HSCs aging. ASP could delay senescence HSCs aging which maybe partly ascribed to the inhibition of oxidative damage and the downregulation of p16 mRNA expression.</p>
Subject(s)
Animals , Female , Male , Mice , Aging , Radiation Effects , Angelica sinensis , Chemistry , Cell Cycle , Radiation Effects , Cells, Cultured , Cellular Senescence , Radiation Effects , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Flow Cytometry , Gene Expression , Radiation Effects , Hematopoietic Stem Cells , Metabolism , Radiation Effects , Mice, Inbred C57BL , Oxidative Stress , Radiation Effects , Polysaccharides , Pharmacology , Random Allocation , Reactive Oxygen Species , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , X-Rays , beta-Galactosidase , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of Angelica sinensis polysaccharides (ASP) on the length of telomere, the activity of telomerase and the expression of P53 protein in mice hematopoietic stem cells (HSCs), and explore ASP's potential mechanism for regulating HSC aging.</p><p><b>METHOD</b>C57BL/6J mice were randomly divided into the normal group, the aging group and the intervention group. The aging group was radiated with X ray to establish the mice aging HSC model. The intervention group was orally administered with ASP during X-ray irradiation, while the normal group was orally administered with NS. Their HSCs were isolated by immunomagnetic beads. Cell cycles analysis and senescence-associated beta-galactosidase (SA-beta-Gal) staining were used to detect changes in aging HSCs. The expression of P53 was determined by western blot analysis. The length of telomere and the vitality of telomerase were analyzed by southern blot and TRAP-PCR, respectively.</p><p><b>RESULT</b>Compared with the normal group, X-ray irradiation could significantly increase the cell ratio of in HSC G1 stage, rate of SA-beta-Gal positive cells and expression of P53 protein, and reduce the length of telomere and the vitality of telomerase. Compared with the aging group, ASP could significantly inhibit the cell ratio of in HSC G1 stage and the increase in the number of SA-beta-Gal positive cells, down-regulate the expression of P53 protein, and increase the length of telomere and the vitality of telomerase in HSCs.</p><p><b>CONCLUSION</b>ASP could antagonize X-ray-induced aging of HSCs, which may be related to the increase in the length of telomere and the activity of telomerase, as well as the down-regulation of the expression of P53 protein.</p>
Subject(s)
Animals , Female , Male , Mice , Angelica sinensis , Chemistry , Cell Cycle , Physiology , Cellular Senescence , Physiology , Hematopoietic Stem Cells , Cell Biology , Metabolism , Mice, Inbred C57BL , Plant Extracts , Chemistry , Pharmacology , Plants, Medicinal , Chemistry , Polysaccharides , Pharmacology , Telomerase , Metabolism , Telomere , Metabolism , Tumor Suppressor Protein p53 , MetabolismABSTRACT
<p><b>AIM</b>To analyze the distribution of single nucleotide polymorphisms in Han ethnic population from Northern China.</p><p><b>METHODS</b>Allele frequencies in a sample of healthy Chinese Hans (n = 204) were determined by polymerase chain reactions followed by restriction analyses with specific endonucleases.</p><p><b>RESULTS</b>The SNP 27916722 A/C in exon 11 from the NCBI database was not detected in this population. And there were significant differences between the allele frequencies of the SNPs (27930097 C/G and 27920978 C/T) in Han ethnic population from Northern China and those in the NCBI.</p><p><b>CONCLUSION</b>It is suggested that the SNPs of sTnT are different in different ethnic populations.</p>