A new form of familial platelet disorder caused by germline mutations in RUNX1 in a pedigree / 中华内科杂志

Objective: To investigate the clinical and biological characteristics of familial platelet disorder (FPD) with germline Runt-related transcription factor (RUNX) 1 mutations. Methods: Patients diagnosed with myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML) with RUNX1 mutations from February 2016 to December 2021 in Wuhan No.1 Hospital underwent pedigree analysis and were screened for gene mutations (somatic and germline). Patients diagnosed with FPD with germline RUNX1 mutations were enrolled and evaluated in terms of clinical characteristics and biological evolution. Bioinformatics analysis was used to assess the pathogenicity of mutations and to analyze the effect of mutated genes on the function of the corresponding protein. Results: Germline RUNX1 mutations were detected in three out of 34 patients suffering from MDS/AML who had RUNX1 mutations. A pedigree of FPD with RUNX1 (RUNX1-FPD) c.562A>C and RUNX1 c.1415T>C mutations was diagnosed, and the mutations were of patrilineal origin. Bioinformatics analysis indicated that the locus at positions 188 and 472 in the AML-1G type of RUNX1 was highly conserved across different species, and that variations might influence functions of the proteins. The mutations were evaluated to be highly pathogenic. Of the nine cases with germline RUNX1 mutations: two patients died due AML progression; one case with AML survived without leukemia after transplantation of hemopoietic stem cells; four patients showed mild-to-moderate thrombocytopenia; two cases had no thrombocytopenia. During the disease course of the proband and her son, mutations in RUNX1, NRAS and/or CEBPA and KIT appeared in succession, and expression of cluster of differentiation-7 on tumor cells was enhanced gradually. None of the gene mutations correlated with the tumor were detected in the four cases not suffering from MDS/AML, and they survived until the end of follow-up. Conclusions: RUNX1-FPD was rare. The mutations c.562A>C and c.1415T>C of RUNX1 could be the disease-causing genes for the family with RUNX1-FPD, and these mutations could promote malignant transformation. Biological monitoring should be carried out regularly to aid early intervention for family members with RUNX1-FPD.

Esophageal Angiolipoma:Report of One Case and Literature Review / 中国医学科学院学报

Esophageal angiolipoma is a rare disease with unspecific clinical manifestations.This paper reported a case of esophageal angiolipoma confirmed by upper gastrointestinal endoscopy and summarized the clinical manifestations,endoscopic and pathological features,treatment and prognosis of the patients by reviewing the relevant literature,aiming to provide references for clinical diagnosis and treatment of this disease in the future.

Changes of intestinal flora in children with acute lymphoblastic leukemia before and after chemotherapy / 中国当代儿科杂志

OBJECTIVES@#To examine the changes of intestinal flora in children newly diagnosed with acute lymphoblastic leukemia (ALL) and the influence of chemotherapy on intestinal flora.@*METHODS@#Fecal samples were collected from 40 children newly diagnosed with ALL before chemotherapy and at 2 weeks, 1 month, and 2 months after chemotherapy. Ten healthy children served as the control group. 16S rDNA sequencing and analysis were performed to compare the differences in intestinal flora between the ALL and control groups and children with ALL before and after chemotherapy.@*RESULTS@#The ALL group had a significant reduction in the abundance of intestinal flora at 1 and 2 months after chemotherapy, with a significant reduction compared with the control group (P<0.05). Compared with the control group, the ALL group had a significant reduction in the diversity of intestinal flora before and after chemotherapy (P<0.05). At the phylum level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Actinobacteria at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05) and a significant increase in the relative abundance of Proteobacteria at 1 and 2 months after chemotherapy (P<0.05). At the genus level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Bifidobacterium at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05); the relative abundance of Klebsiella in the ALL group was significantly higher than that in the control group at 1 and 2 months after chemotherapy and showed a significant increase at 1 month after chemotherapy (P<0.05); the relative abundance of Faecalibacterium in the ALL group was significantly lower than that in the control group before and after chemotherapy and showed a significant reduction at 2 weeks and 1 month after chemotherapy (P<0.05). The relative abundance of Enterococcus increased significantly at 1 and 2 months after chemotherapy in the ALL group (P<0.05), and was significantly higher than that in the control group (P<0.05).@*CONCLUSIONS@#The diversity of intestinal flora in children with ALL is significantly lower than that in healthy children. Chemotherapy significantly reduces the abundance of intestinal flora and can reduce the abundance of some probiotic bacteria (Bifidobacterium and Faecalibacterium) and increase the abundance of pathogenic bacteria (Klebsiella and Enterococcus) in children with ALL.

Therapeutic efficacy observation on moxibustion with moxa of different storage years for moderate-to-severe primary knee osteoarthritis / 针灸推拿医学（英文版）

Objective: By observing the differences in Western Ontario and McMaster Universities osteoarthritis index (WOMAC) and visual analog scale (VAS) scores in moxibustion treatment for moderate-to-severe primary knee osteoarthritis (KOA) with moxa of different storage years (3-year moxa and 1-year moxa from Qichun, Huanggang City, Hubei Province, China) through a randomized clinical trial, to objectively evaluate the differences in therapeutic efficacy of moxibustion with moxa of different storage years. Methods: A total of 63 patients with moderate-to-severe KOA who met the inclusion criteria were randomized into moxibustion group 1 and moxibustion group 2 by central randomization method, with 32 cases in moxibustion group 1 and 31 cases in moxibustion group 2. Moxibustion group 1 was treated with moxa stored for 3 years, and moxibustion group 2 was treated with moxa stored for 1 year. Dubi (ST 35), Neixiyan (EX-LE 4) and Heding (EX-LE 2) were selected in both groups, and the treatment lasted 20 min per time, 3 times a week. The immediate efficacy was compared after 6 times of treatment, and long-term efficacy was compared at follow-up 4 weeks after the end of treatment. Results: During the treatment, there were 2 dropouts in moxibustion group 1, and 1 dropout in moxibustion group 2. The total effective rate in the two groups was 83.3％ and 60.0％, respectively. Followed up at 4 weeks after the end of treatment, the total effective rate in the two groups was 80.0％ and 66.7％, respectively. There were no statistical differences between the two groups (both P>0.05). After treatment and 4 weeks after the end of treatment, the WOMAC and VAS scores in both groups decreased significantly compared with those before treatment (all P<0.01); the scores of stiffness item of WOMAC in moxibustion group 1 were lower than those in moxibustion group 2 (both P<0.05); there were no statistical differences in the scores of pain item and dysfunction item of WOMAC, and VAS scores between the two groups (all P>0.05). Conclusion: Moxibustion with moxa of different storage years (stored for 3 years and 1 year) both can improve the pain, stiffness and motor function in patients with moderate-to-severe KOA. While moxa stored for 3 years has a better therapeutic efficacy in improving stiffness of the knee joint than that stored for 1 year.

The relationship between serum Golgi protein 73 and both liver inflammation and fibrosis in patients with chronic hepatitis B / 解放军医学杂志

Objective To explore the correlation between serum Golgi protein 73(GP73) and both liver inflammation and fibrosis in patients with chronic hepatitis B(CHB). Methods Altogether 322 patients who underwent liver biopsy were enrolled in this retrospective study, and GP73, ALT, AST, LSM, FIB-4, APRI were analyzed. The relationship between serum GP73 and both liver inflammation and fibrosis was analyzed. Univariate and multivariate logistic regression analysis were used to evaluate the predictive value of GP73 for significant liver injury. Results 322 CHB patients ranged from 18 to 66 years old, including 231 males (71.7%) and 91 females (28.3%), and the median of GP73 was 65.6(35.8-129.6) ng/ml. The serum level of GP73 stepwise increased with increase of liver inflammation grade and liver fibrosis stage, and GP73 was significantly correlated with liver inflammation grade (?=0.536, P<0.001) and fibrosis stage (?=0.536, P<0.001). The degree of liver inflammation or fibrosis at the same stage was analyzed and the results indicated that the serum GP73 levels were significantly correlated with the severity of liver inflammation, while the association between serum GP73 and the stage of liver fibrosis was weaker. In term of the advanced liver inflammation or fibrosis stage, univariate and multivariate analysis revealed that GP73 was simultaneously determined as an independent risk factor for predicting liver inflammation (P<0.001) and fibrosis (P<0.05). Conclusion Serum GP73 is well correlated with liver inflammation and fibrosis in CHB patients, and it might be a new biomarker to predict liver inflammation and fibrosis.

Ultracentrifugation, ExoPerfectTM-MU and PEG6000 for the extraction and identification of seminal plasma exosomes: A comparative study / 医学研究生学报

Objective　No studies have been reported on the comparison of ultracentrifugation, ExoPerfectTM-MU and PEG6000 in extracting seminal plasma exosomes. This article aimed to compare the three methods for the extraction and identification of seminal plasma exosomes. Methods　Semen samples were obtained from 30 healthy donors and randomly divided into three portions, followed by extraction of exosomes from the seminal plasma by ultracentrifugation, ExoPerfectTM-MU, and 8%PEG6000, respectively. The size of the extracted exosomes was measured with the nanoparticle tracking analyzer (NTA), their morphology observed under the transmission electron microscope (TEM), and their protein biomarkers detected by Western blot.　Results　Significantly higher expressions of CD63 and TSG101 were found in the exosomes extracted by ultracentrifugation than in those extracted by ExoPerfectTM-MU and 8%PEG6000 (P0.05). Compared with the 8%PEG6000 group, the ultracentrifugation and ExoPerfectTM-MU groups showed significantly higher concentrations ([11.90±1.78] vs [21.20±0.98] and [19.74±1.45]×108/mL, P<0.01) and numbers of seminal plasma exosomes under TEM (4.7±1.7 vs 7.0±1.6 and 6.0±1.6, P< 0.01).　Conclusion　Ultracentrifugation, ExoPerfectTM-MU and 8%PEG6000 are all capable of successful extraction and identification of seminal plasma exosomes, but the former two yield more exosomes, the latter one gives a higher purity, and ExoPerfectTM-MU is simple and convenient in operation.

Clinical value of HE4,CA125 and ROMA in diagnosis of ovarian epithelial malignant tumor / 中国实用妇科与产科杂志

OBJECTIVE: To investigate the value of HE4,CA125 and ovarian malignant tumor risk prediction models(ROMA)in the diagnosis of ovarian epithelial malignant tumors.METHODS: The clinical and pathological data of 247 patients with ovarian tumors(EOC 139 cases,BOT 18 cases,and benign ovarian tumor 90 cases)and 39 patients with uterine fibroids in Liaoning Tumor Hospital from September 2016 to August 2017 were retrospectively analyzed. The levels of serum CA125 and HE4 were measured before operation. The ROMA values were calculated and the relationship between CA125,HE4,ROMA values and clinical pathological parameters were analyzed. The diagnostic evaluation index was calculated,the receiver operating characteristic(ROC)curve was drawn,and the AUC value was also calculated.RESULTS: The positive rate of HE4 in the ovarian epithelial malignant tumor group was significantly higher than that in other groups before and after menopause,the difference being statistically significant(P0.05).There was no significant statistical difference in the positive rate of ROMA before or after menopause(P>0.05).The sensitivity of CA125 was higher than that of HE4 and ROMA. Specificity of HE4 was higher than that of CA125 and ROMA.Correct diagnosis index of ROMA was higher than that of HE4 and CA125.CONCLUSION: For the diagnosis of ovarian malignant epithelial tumors,the combined detection of serum HE4 and CA125 and ROMA model is superior to the individual detection of HE4 and CA125.

Early diagnosis of acute kidney injury in aged patients undergoing percutaneous coronary intervention / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

In aged patients, acute kidney injury (AKI) is a common clinical complication after percutaneous coronary intervention (PCI), highlighting the need for timely and certain diagnosis of this disease. A single centre, nested case-control study was conducted, which assessed the usefulness of urinary liver-type fatty acid-binding protein (uL-FABP), neutrophil gelatinase-associated lipocalin (uNGAL), and kidney injury molecule-1 (uKIM-1) for early detection of AKI. One hundred and thirty-two patients at or over 60 years old undergoing PCI were included. Serum creatinine (SCr) was measured before PCI, 24 and 48 h after PCI; uL-FABP, uNGAL, and uKIM-1 were measured before PCI, 6, 24, and 48 h after PCI. We identified 16 AKI patients and selected 32 control patients matched by admission time (<1 week), age (±5 years), and gender. In the receiver operating characteristic (ROC) curve analysis, the areas under the curve (AUCs) for the relative measurements of uL-FABP, uNGAL, and uKIM-1 were 0.809, 0.867, and 0.512 at 6 h after PCI, and 0.888, 0.840, and 0.676 at 24 h after PCI, respectively. AUC for the combination of uL-FABP and uNGAL was 0.899 at 6 h after PCI, and 0.917 at 24 h after PCI. Thus, measurement of uL-FABP and uNGAL levels at 6 and 24 h after PCI may be useful in detecting AKI in aged patients. Measurement of uKIM-1 levels provides inferior predictive power for early diagnosis of AKI.

Changes of T Cell Subsets and Their Relationship with Clinical Features and Prognosis in Patients with Acute Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the relationship between T cell subsets and clinical data.</p><p><b>METHODS</b>mononuclear cells were collected from 103 patients with acute leukemia (AL) and 28 healthy volunteers, and percentage changes of CD3CD4, CD3CD8 and CD4 CD25 Foxp3 cell subsets were assayed by flow cytometory. Relationship between the T subsets and clinical features of the patients was analyzed.</p><p><b>RESULTS</b>Ratio of CD3 T cells decreased more significantly in patients with >50% blast cells than that in patients with <50% blast cells, while the ratio of Treg between the 2 groups was not significantly different. Treg increased more statistically significantly in the patients with CD34 leukemia cell than that with CD34 leukemia cells. In constrast to the relationship between prognosis and immune cells in the patients from 3 groups (low, intermediate and high-risk group) it was found that Treg cells increased more significantly in high-risk group than that in low-risk group. By continuously monitoring immune cells in 18 patients, it was found that Treg cells gradually increased during the first 3 courses of chemotherapy, then began to decreased in the 4th course, finally approached gradually to the normal value in the 6th course, and this change correlated with the clinical remission after chemotherapy. Treg cell number in the patients with AL was significantly higher than that in healthy controls, and Treg cell number during the onset and recurrence was significantly higher than that in the period of complete remission (continuous remission for over 6 months). Compared with the changes of immune cell number between different types of disease, it was found that Treg cells were increased more significantly in acute myeloid leukemia (AML) than that in acute lymphoblastic leukemia (ALL). Proportion of Treg cells, Treg/CD4 decreased more significantly after the 1st course of chemotherapy in the group with complete remission (CR) than that in the group without CR. The complete remission rate and recurrence rate were 68.9% and 20% respectively in the group with >10% Treg cells, while the complete remission rate and recurrence rate were 85.7% and 7.69% respectively in the group with.<10% Treg cells. In comparison of the 6 recurrent patients with 32 patients with sustained CR, it was found that the ratio of Treg cells and Treg/CD4 was increased more significantly in the patients with relapse than that with CR and in control group.</p><p><b>CONCLUSION</b>Dynamic change of Treg cells in the peripheral blood was closely related with clinical feature, recurrence and prognosis in the patients with acute leukemia.</p>

Orf virus ORF047 gene eukaryotic expression and cell localization / 中国人兽共患病学报

The object of study was to clone the gene of ORFV ORF047 and study the eukaryotic expression and cell localization,making the theoretical basis for the subsequest screening of protein that interact with ORF047.ORF047 gene was amplificated by the specifical primer from the DNA of ORFV using PCR,the length was 735 bp,compared with L1 published in NC-005336.1,the homologies of the nucleotide acid sequence and amino acid sequence were 98.8％ and 98.8％.In order to defined the expression and location of the ORF047 gene in cell,the recombinant plasmid pEGFP-ORF047 was constructed and transfected into 293T cell,after 36 h,the green fluorescence could be observed under fluorescence microscope,and 54 kD protein was detected by western bloting.The plasmid of pHcRed1-Nuc,pHcRed1-Mito and pHcRed1-ER with the recombinant plasmid of pEGFP-ORF047 was cotransfected to veroE6 cell respectively,that fusion protein of ORF047 was mainly located in the cytoplasm,a small amount in the mitochondriabyconfocal microscopy analysis.

Clinical study of laser photocoagulation combined with VEGF antagonists for DME / 国际眼科杂志(Guoji Yanke Zazhi)

· AIM:To investigate the influence of laser photocoagulation and vascular endothelial growth factor (VEGF) antagonists used alone or as combination therapy on clinical efficacy and safety of patients with diabetic macular edema (DME).· METHODS:Totally 150 patients (156 eyes) with DME were chosen in the period from October 2014 to October 2016 in our hospital and randomly divided into both group including Group A (50 patients 52 eyes) with laser photocoagulation used alone,Group B (50 patients 51 eyes) with VEGF antagonists used alone and Group C (50 patients 53 eyes) with combination therapy;and the best corrected visual acuity,macular fovea thickness and retinal neovascularization leakage area before and after treatment and the complications incidence of both groups were compared.· RESULTS:The best corrected visual acuity of Group B and Group C in 3,6 and 12mo after treatment were significant better than that of Group A (P＜ 0.05).The macular fovea thickness of Group B and Group C in 3,6 and 12mo after treatment were significant lower than that of Group A (P＜0.05).The retinal neovascularization leakage area of Group B and Group C in 3mo after treatment were significant smaller than that of Group A (P＜0.05).The retinal neovascularization leakage area of Group C in 6 and 12mo after treatment were significant smaller than that of Group A and Group B (P＜0.05).There was no significant difference in the complications incidence among 3 groups (P＞0.05).· CONCLUSION:Laser photocoagulation combined with VEGF antagonists in the treatment of patients with DME can efficiently improve visual acuity,reduce macular foveal thickness,control retinal neovascularization leakage and not increase adverse reactions.

The clinical observation of Compound Xuanju Capsules Combined with methotrexate treating rheumatoid arthritis / 中成药

AIM To evaluate the clinical effects of Compound Xuanju Capsules (CXC,Polyrhachis vicina Roger,Epimedii Folium,Lycii Fructus,Cnidii Fructus) combined with methotrexate (MTX) on patients with moderate or severe rheumatoid arthritis (RA).METHODS One hundred and eighty RA patients were randomized into 3 groups:CXC group,MTX group and combined group (dosed with both CXC and MTX) for a 48-week intervention.The clinical observation on the changes of the signs and symptoms,erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),health assessment questionnaire (HAQ),visual analogue scale (VAS) and disease active score (DAS) 28 were performed before and after each treatment.ACR20,American College of Rheumatology 20％ improvement criteria was taken as the primary end point,and ACR50 and ACR70 as the secondary end points to standardize the patients' response measurement,and all adverse reactions were recorded as well.RESULTS At the time point right after the 12th week,the ACR20 response rate of the combined group (41.5％) was significantly higher than that of the CXC group (19.6％,P ＜ 0.05) and the MTX group (24.1％,P＜0.05).The respective ACR50 (2.1％) and ACR70 (20.8％) response rate of the combined group were significantly higher than those of CXC group and MTX group (P ＜ 0.05).At the time point right after the 24th week,the combined group still demonstrated its significantly higher ACR20 response rate (81.1％) to the CXC group (30.4％,P ＜ 0.05) and the MTX group (68.5％,P ＜ 0.05).The similar superiority in ACRS0 (60.4％) and ACR70 (54.7％) response rate of the combined group to the CXC group and the MTX group were found (P ＜0.05).After the 48th week,the combined group displayed its significantly higher ACR70 response rate (75.7％) to CXC group and MTX group (P ＜ 0.05).Given the reduction of DAS28,HAQ and VAS from the12thweek's5.26±0.83),(22.2±10.3),(6.0±0.4) to the 24thweek's (4.21 ±0.91),(17.1±10.3),(2.4±2.2),andthe48thweek's (2.19±0.56),(10.4±5.0),(0.8±0.9),the combined group's more outstanding performance compared to CXC group and MTX group started right after the 24th week (P ＜ 0.05).Generally,no difference in adverse events was detected between the combined group and MTX group (P ＞ 0.05).CONCLUSION The combined use of CXC and MTX can be an effective and safe treatment for moderate and severe rheumatoid arthritis.

S100A4, a potential therapeutic target on bladder cancer stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of S100A4 gene silencing mediated by small interfering RNA (siRNA) on the proliferation of bladder cancer stem cells (CSCs) and their capacity of xenograft tumor formation.</p><p><b>METHODS</b>MB49 bladder cancer stem cells (MCSCs) were isolated and identified. The differentially expressed protein S100A4 was identified in MCSCs using isobaric tags for relative and absolute quantitation technology (iTRAQ). A siRNA targeting S100A4 was constructed and transfected into MCSCs, and its inhibitory effects on S100A4 expression in MCSCs were assessed with Western blotting and qPCR. The effects of siRNA-mediated S100A4 silencing on the proliferation and xenograft tumor formation ability of MCSCs were observed.</p><p><b>RESULTS</b>Among the 65 differentially expressed proteins identified by iTRAQ combined with LC/MS/MS, S100A4 protein showed the most distinct differential expression in MCSCs. Transfection of MCSCs with S100A siRNA significantly inhibited the expressions of S100A4 at both mRNA and protein levels, caused obvious suppression of the cell proliferation, and attenuated the xenograft tumor formation ability of the cells in nude mice.</p><p><b>CONCLUSION</b>S100A4 in MCSCs is associated with the recurrence and metastasis of bladder cancer. S100A4 may serve as a potential therapeutic target for eliminating bladder CSCs.</p>

Establishment of an individualized three-dimensional finite element model of type A coronary artery lesions / 中国组织工程研究

BACKGROUND:A cardiac model can be established by finite element analysis based on patient's MRI imaging data.The established model can be used to evaluate the rheological changes of the coronary artery by liquid-solid coupling.OBJECTIVE:To establish finite element models of the heart and coronary artery in patients with type A coronary artery disease using finite element analysis software,followed by three-dimensional (3D) printing,thereby providing a scientific basis for further simulation of interventional surgery.METHODS:Three patients with type A coronary artery lesions underwent MRI scanning from the aortic arch to the apex.The MRI images were then imported into the Mimics 17.0 software in Dicom format,and a complete cardiac model involving the coronary arteries was established by modeling and geometry cleanup.The 3D model was imported into Geomagic Studio 11.0 software,and was further processed.Finally,the 3D model was imported into ANSYS14.0 finite element analysis software.The finite element model with biofunction was established by attaching the material properties,followed by 3D printing on a 3D printer.RESULTS AND CONCLUSION:The 3D finite element model of type A coronary artery lesion was established successfully in three cases.The established heart model in each case presented with grid-based hexahedral solid elements.The number of solid elements was 24 532,25 771,and 24 330,respectively.In the meanwhile,the model of each coronary branch was established:the number of element at the right coronary artery was 3 320,3 518,and 3 310;the number of elements at the circumflex branch was 1 148,1 176,and 1 164;and the number of elements at the anterior descending coronary artery was 1 025,1 049,and 1 068,respectively.Afterwards,the 3D printing was performed successfully.These results suggest that the established 3D finite element model of the heart with coronary arteries,after 3D printing,displays the right coronary artery,anterior descending artery,circumflex artery and coronary sinus clearly,which paves ways for interventional simulation.Most importantly,it lays a solid foundation for the study on the blood-vessel dual-directional coupling,which is expected to be a new scientific method for rheological research.

Pro-apoptotic effect of a tumor suppressor gene WWOX on ovarian cancer stem cells and its mechanism / 中国组织工程研究

BACKGROUND:WWOX,a tumor suppressor gene,can affect the growth of ovarian cancer stem cells;however,there is no report on whether its mechanism of action is related to Hedgehog signaling pathway.OBJECTIVE:To investigate the effect and mechanism of overexpression of WWOX on the apoptosis of ovarian cancer stem cells.METHODS:pcDNA3.1-WWOX (pcDNA3.1-WWOX group) and pcDNA4.0-WWOX (pcDNA4.0-WWOX group) were transferred into ovarian cancer stem cells,respectively;and meanwhile,pcDNA3.1 (pcDNA3.1 group) and pcDNA4.0 (pcDNA4.0 group) were transferred into the cells.A non-transfection group (only with Lipofectamine2000) was set up.After cultured 48 hours,the levels of WWOX in the pcDNA3.1-WWOX group and pcDNA4.0-WWOX group were detected using western blot assay,and the cell proliferation and apoptosis were detected using MTT assay and flow cytometry,respectively.Western blot assay was also used to detect the levels of Hedgehog signaling pathway associated proteins,SHH,PTCH1,Gli-1,SMO and apoptosis-related protein Cleaved Caspase-3 in the cells.Cyclopamine,Hedgehog signaling pathway inhibitor,was used in ovarian cancer stem cells without transfection (cyclopamine group) and after the transfection of WWOX overexpression vector (WWOX+cyclopamine group) followed by 48 hours of culture,and then MTT,flow cytometry and western blot detections were performed.RESULTS AND CONCLUSION:(1) The expression level of WWOX in the pcDNA4.0-WWOX group was significantly higher than that in the pcDNA3.1-WWOX group (t=27.84,P=0.00).The ovarian cancer stem cells which were transfected with pcDNA4.0-WWOX were used to overexpress WWOX in the late experiment.(2) Overexpression of WWOX could inhibit the proliferation of ovarian cancer stem cells and promote the apoptosis of ovarian cancer stem cells.(3) Overexpression of WWOX could inhibit the expression of Gii-1,PTCH1,SMO and SHH in ovarian cancer stem cells,and promote the expression of Cleaved Caspase-3.(4) Cyclopamine could inhibit the expression of SHH,PTCH 1,Gli-1,SMO,and promote the expression of Cleaved Caspase-3.Cyclopamine had obvious inhibitory effect on Hedgehog signaling pathway.(5) Cyclopamine could enhance the apoptosis induced by overexpression of WWOX in ovarian cancer stem cells,and enhance the inhibition of proliferation of ovarian cancer stem cells induced by overexpression of WWOX.To conclude,WWOX effects on proliferation and apoptosis of ovarian cancer stem cells may be related to the inhibition of Hedgehog signaling pathway.

Changes of MMP-9 expression in rabbit retina after laser irradiation at different thresholds / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the changes of MMP-9 expression in rabbit retina after laser irradiation at different thresholds of 577nm.METHODS: Twenty-six pigmentation rabbits were randomly divided into normal control group(n=2), conventional photocoagulation group (n=6) and subliminal micropulse laser photocoagulation group (n=18).The conventional photocoagulation group was treated with 577nm laser photocoagulation, subcutaneous micro-pulsed laser photocoagulation at a working loading rate of 9%, 12% and 15%, respectively.Eighteen rabbits were again divided into three subgroups according different powers of subthreshold working loading rate of 9%(n=6), 12%(n=6) and 15%(n=6) that undertook, respectively.The expression of MMP-9 on the retina of rabbit eyes was detected by immunohistochemistry.RESULTS: In the conventional photocoagulation group, the expression of MMP-9 in the RPE layer and the visual cell layer was strongly positive, which was significantly higher than that in the sub-micro pulse group(P0.05).CONCLUSION: The 577nm subliminal micro-pulsed photocoagulation has high selectivity to retinal pigment epithelium at working load rate of 9%, 12% and 15%, and no damage to retinal nerve fiber layer, which is safer than conventional 577nm laser photocoagulation.

Mechanisms of Lung Cancer Caused By Cooking Fumes Exposure: A Minor Review / 中国医学科学杂志(英文版)

Cooking fumes (CFs) are mixtures of many toxic components, such as aldehydes, heterocyclic amines, polycyclic aromatic hydrocarbons, fat aerosols and particulate matters. CFs exposure has been proven to be associated with many diseases. Lung cancer takes the leading place among the diseases being reported caused by CFs exposure. Molecular and biochemical studies have found that CFs exposure may lead to lung cancer by gene damage, formation of reactive oxygen species, blockage of related proteins' function, and even cell death. However, reviews about the mechanisms of how CFs exposure leads to lung cancer are still lacking. Elucidation of the mechanisms of lung cancer caused by CFs exposure may provide a new insight into the prevention of lung cancer caused by CFs exposure, as well as laying the foundation for the toxicity study of CFs. In this minor review, the mechanisms of how CFs exposure leads to lung cancer were summarized and discussed.

Extraction and identification of semen-derived exosomes using PEG6000 / 南方医科大学学报

<p><b>OBJECTIVE</b>To extract and identify semen-derived exosome using PEG6000.</p><p><b>METHODS</b>Exosomes were extracted from semen specimens from 6 healthy volunteers with step-by-step centrifugations and ultracentrifugation prior to 8% PEG6000 enrichment. The extracted exosomes were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blotting.</p><p><b>RESULTS</b>The pellets obtained were round or elliptic membrane vesicles 30 to 150 nm in diameter with intact double membranes and contained low electron density material. The pellets expressed CD63, ALIX and TSG101 molecules but not calnexin that was expressed in sperm cells.</p><p><b>CONCLUSION</b>The PEG6000-based method for extraction of exosomes from semen samples facilitates future studies of seminal exosomes.</p>

In vitro metabolism of anti-tumor compound E7 in different species of liver microsomes / 中国中药杂志

To investigate the metabolic stability of E7 in liver microsomes of human, Beagle dog, Cynomolgus monkey and SD rats, and compare the metabolic differences between different species. Selective chemical inhibitors were used to determine the effects of different inhibitors on E7 metabolic rate, and predict the main enzymes involved in E7 metabolism in rat liver microsomes. The experimental results showed that the in vitro half-lives (T1/2) of E7 in liver microsomes of human, dog, monkey and rats were 57.75, 69.30, 16.90,30.13 min respectively. Their intrinsic clearance rate was 0.004 8, 0.004 0, 0.016 4 and 0.009 2 mL•min⁻¹•mg⁻¹ respectively. Hence, it could be speculated that the metabolic rate of E7 was similarly slow in human and dog liver microsomes; while it was similarly fast in monkey and rat liver microsomes. There was significant difference in metabolic rate of E7 between different species. The results showed that CYP2E1, CYP2A6, CYP1A2 and CYP2D6 might participate in metabolism of E7, while the contribution of polymorphic CYP3A4 was small.

Survival analysis of osteosarcoma patients with pulmonary metastasis undergoing different therapeutic methods / 肿瘤

Objective: To analyze the relationship between the prognosis and the different therapeutic methods in osteosarcoma patients with pulmonary metastasis. Methods: The clinical data of 87 osteosarcoma patients with pulmonary metastasis admitted in Sixth People's Hospital from January 2006 to August 2011 were retrospectively reviewed. All patients were followed up. According to the different therapeutic methods for the metastatic lung lesions, these 87 patients were divided into three groups: surgery combined with chemotherapy group (21 cases), gamma-knife radiosurgery combined with chemotherapy group (26 cases), and chemotherapy alone group (40 cases). Kaplan-Meier method was used for survival analysis. The prognosis-related factors were analyzed by univariate and multivariate analyses. Results: The median progression-free survival (PFS) of patients in surgery combined with chemotherapy group or gamma-knife radiosurgery combined with chemotherapy group (8 months and 6 months) was longer than that in the chemotherapy alone group (3 months) (both P < 0.01). Three-year survival rates of osteosarcoma patients in surgery combined with chemotherapy group, gamma-knife radiosurgery combined with chemotherapy group, and the chemotherapy alone group were 38.10%, 30.77% and 12.50%, respectively. The unilateral or bilateral lung metastasis, initial treatment or retreatment, the number and distribution type of lung metastatic lesions, metastases of other sites (not lung), and the therapeutic methods for the lung lesions were correlated with prognosis (P < 0.05). The therapeutic method for lung metastatic lesions was an independent prognostic factor (P < 0.05). Conclusion: Chemotherapy combined with surgery or gamma-knife radiosurgery can effectively improve the survival of osteosarcoma patients with lung metastasis.