Marine actinomyces LYG-1 was isolated from marine mud flats in Lianyungang,China.Strain LYG-1 was identified using the methods of morphology,physiological and ehemotaxonomic characterization and 16S rRNA gene sequence analysis.The results showed that strain LYG-1 was a marine variable species of Streptomyces roseosporus.The fermentation broth of strain LYG-1 exhibited conspicuous antitumor activity against HepG2,MCF-7,HCT116 and MDA-MB-231 cell lines,and the IC50 values were defined by MTT method respectively.

Objective To investigate the effects of human amniotic membrane on the survival of the crossboundary perforator flap in rats through distal subcutaneous implantation.Methods From February,2016 to December,2016,38 SD rats weighing 270-300 g were randomly divided into experimental group and control group (n=19).A three-territory perforator flap (3 cm× 10 cm) with the iliac artery as the pedicel elevated on the right dorsum of the rats.Experiment group,a section of amnion was sutured to the distal subcutaneous of the flap and then the flap was sutured back to its donor site.Control group,after the flap elevated,directly sutured back to its donor site.The blood flow of the pedicel were detected by laser Doppler flowmetry at the day before surgery,1 day after surgery,3 and 7 days after surgery,with 3 rats in every group.When the rats were anesthetized,biopsies were taken from the choke area Ⅱ at the day before surgery,3 and 7 days after surgery,with 9 rats in every group.When the rats were anesthetized,HE was used to compare the diameter size of the artery and vein at the same site.At day 7,measure the survival area of the flap,and 3 of them observe the vessel of the flap by lead oxide-gelatine technique.Results The survival rate of the experimental group and the control group after 7 days were (89.09±4.23)％ and (74.56-±5.59)％ respectively,the experimental group was significantly higher than that of the control group (t=5.48,P=0.00).X-ray showed that 7 days after operation,the pedicel of iliac artery in the experimental group was bigger than that in the control group.The blood flow detection showed that the blood flow of the experimental group was higher than that of the control group (t=2.39,3.06;P=0.03,0.00,respectively) at day 1 and day 3 after surgery.Hematoxylin eosin staining showed that arteriovenous tube diameter at the Choke Ⅱ were gradually increased on the day3 and day7 after surgery in both group,but the experimental group diameter expanded multiple was higher than that of control group (t=3.52,3.50;P=0.02,0.02,respectively).Conclusion Human Amnion subcutaneous embedding may improve the blood flow of the vascular pedicle,expand the microvascular at Choke area Ⅱ,improve microcirculation,as a result to promote the survival of cross perforator flap,while the mechanism is needed to understanding.

Objective To investigate the effects of DMOG on the microcirculation of the choke-area and the survival of the cross-boundary flap in rats via tail vein injection.Methods Rats with ischemic three-territory perforator flaps on the dorsum were treated with DMOG at a dosage of 40 mg/kg body weight via tail vein injection at 1 day before surgery(day-1),the time of surgery(day 0),1 day after surgery(day 1),2 days after surgery(day 2) and 3 days after surgery(day 3).Control group received sterile saline at the same time points and same dosage via tail vein injection.① Draw materials from the choke-area at day 1,day 3 and day 7,HE stain was used to compare the diameter size of the artery and vein at the same site.② Western blotting to check the expression of PCNA and HIF-1α,ELISA to detect the content of PCNA,HIF-1α,SDF-1α and VEGF at day 7.③At day 7,measure the survival area of the flap and observe the vessel of the flap by lead oxide-gelatine technique.Results ① There was a greater survival rate of (96.3 ± 5.1)％ in the treatment group than in the control group with (73.9 ± 5.8)％ at day 7 (P ＜ 0.05).② The diameter size of the arterioles and venules were dilated in both groups until postoperative days 7.But the treatment group was more expanded than the control group at day 3(2.20 ± 0.26 vs.1.50 ± 0.20,P ＜ 0.05) and day 7(3.67 ± 0.35 vs.2.03 ± 0.15,P ＜ 0.05).③ The skin expression of PCNA and HIF-1α in the treatment group were greater than the control group(P ＜ 0.05) at day 7.④ The content of skin PCNA in the treatment group and control group were(8.95 ± 0.71) ng/mg and (4.15 ± 0.72) ng/mg,HIF-1α were(5.04 ± 0.50)ng/mg and (2.98 ± 0.29) ng/mg,SDF-1α were (2.91 ± 0.61) ng/mg and (1.39 ± 0.62) ng/mg,and VEGF were(2.17 ± 0.41) ng/mg and (0.95 ± 0.44) ng/mg,respectively.The treatment group was greater than the control group (P ＜ 0.05).Conclusion DMOG can improve the microcirculation of the choke area,and then increase the survival of the perforator skin flaps in rats via tail vein injection.

Objective:To investigate the urineinterleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-33 (IL-33) and matrix metalloproteinase-9 (MMP-9) in patients with IgA nephropathy (IgAN) and their predictive value for disease progression.Methods:110 IgAN patients admitted to Zhuozhou Hospital from January 2018 to October 2021 were selected and divided into IgAN progression group (39 cases) and IgAN non progression group (71 cases) according to the progress of IgAN patients. According to the Oxford Classification Standard System (MEST-C) of IgAN, they were divided into MEST-C≥3 group (42 cases) and MEST-C<3 group (68 cases). According to the estimated glomerular filtration rate (eGFR), they were divided into eGFR≥50 ml/(min·1.73 m 2) group (group A, 63 cases) and eGFR<50 ml/(min·1.73 m 2) group (group B, 47 cases). According to the amount of urinary protein, they were divided into urinary protein≥1.5 g/24 h group (66 cases) and urinary protein<1.5 g/24 h group (44 cases). Multivariate logistic regression was used to analyze the risk factors affecting the progression of IgAN. Receiver operating characteristic (ROC) curve was drawn to analyze the value of urinary IL-6, IL-8, IL-33 and MMP-9 levels in predicting the progression of IgAN. Results:The urinary IL-6, IL-8, IL-33 and MMP-9 levels in IgAN progression group were significantly higher than those in IgAN non progression group, while the serum albumin, eGFR and complement C3 in the IgAN progression group were lower than those in the IgAN non progression group (all P<0.05). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the MEST-C≥3 group were significantly higher than those in the MEST-C<3 group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 levels in the eGFR<50 ml/(min·1.73 m 2) group were significantly higher than those in the eGFR≥50 ml/(min·1.73 m 2) group (all P<0.001). The urinary IL-6, IL-8, IL-33 and MMP-9 in the urinary protein≥1.5 g/24 h group were significantly higher than those in the urinary protein<1.5 g/24 h group (all P<0.001). Multivariate logistic regression analysis showed that the course of disease, serum albumin, eGFR, urinary protein, IL-6, IL-8, IL-33 and MMP-9 were risk factors affecting the progression of IgAN (all P<0.005). The ROC curve showed that the area under the curve (AUC) of IL-6, IL-8, IL-33 and MMP-9 in predicting the progression of IgAN was 0.956 (95% CI: 0.891-0.998), and the sensitivity and specificity were 98.4% and 86.2%, respectively. Conclusions:The elevated levels of urinary IL-6, IL-8, IL-33 and MMP-9 are closely related to the progress of IgAN, and the combination of these four indicators has a good value in predicting the progress of IgAN.

Objective In this study,a multiplex PCR amplification system was constructed based on fluorescent labeling PCR and LDR,to provide a new strategy for analyzing severely degraded DNA.Methods Eight SNP loci (rs10802248,rs10516197,rs10488372,rs2278945,rs4757318,rs4887255,rs4889002,and rs9304473) were selected.Their LDR probes and PCR primers of linked products were designed and synthesized.Ligase detection reaction,PCR amplification,and capillary gel electrophoresis (CEG) were performed to establish the multiplex LDR-PCR amplification system.Results The genotypes of these 8 loci were obtained simultaneously by the fluorescence-labeled multiplex LDR-PCR amplification method.The loci profiles obtained by fluorescence-labeled multiplex LDR-PCR amplification were in accordance with those obtained by direct sequencing of the polymorphic regions in samples from all individuals.By fluorescence-labeled multiplex LDR-PCR amplification,the 8 SNP loci were efficiently amplified from the severely degraded FFPET DNA.Conclusion Eight SNP loci results could be obtained simultaneously by using the multiplex LDR-PCR amplification system,which is a simple,efficient,and practical SNP genotyping method with accurate and reliable results for highly degraded samples.

Objective: To investigate hepatitis C virus(HCV)genotyping and the serum HCV-RNA concentration in patients infected with different HCV genotypes and to provide information for evaluation of disease condition and anti-viral treatment efficacy. Methods: A total of 60 anti-HCV positive serum samples were collected before antiviral treatment. RT-PCR was performed for the 5′ non-cording region and was followed by nucleotide sequencing for HCV genotyping. Meanwhile, serum HCV-RNA concentration was detected by quantitative PCR. SPSS21.0 and Graphpad Prism 5.0 software were used for data analysis. Analysis of variance (ANOVA) was used for comparison among multi-groups and the t-test was used for comparison between two groups. Results: The frequencies of HCV genotypes 1b, 3a, 1a and 2a were 48.3% (29/60), 23.3% (14/60), 16.7% (10/60) and 10% (6/60), respectively. And, there is one subtype 2c was detected in this study. The mean serum viral concentration with standard deviation of HCV in genotype 1a, 1b, 2a, and 3 a were 5.46±1.19, 6.22±0.78, 5.47±0.65, and 5.38±0.98 log₁₀ (IU/ml) respectively. Conclusions The infection rate of HCV genotype 1 was significantly higher than that of genotype 2 and 3 (P<0.01). The statistical analysis showed the serum HCV-RNA concentration in patients with subtype 1b was significantly higher than that of the subtype group 1 a, 2 a, and 3 a (P<0.05). The study of the relationship between HCV genotypes and the serum HCV-RNA concentration may contribute to anti-viral treatment prescription for hepatitis C patients.

Objective:To describe the clinical features and prognosis of congenital anomalous origin of coronary artery(AOCA) in children to increase our understanding of the disease.Methods:This retrospective study included children diagnosed with AOCA using computed tomography coronary angiography(CTCA) admitted to the Department of Cardiology, Beijing Children′s Hospital, Capital Medical University, from January 1, 2014 to December 31, 2019.The clinical presentations, laboratory results, imaging analyses, treatments, and prognoses of these patients were analyzed.Results:A total of 208 children, including 105 boys and 103 girls, we evaluated the ages(9.03±4.18)years old with AOCA.Of these, 157 cases(75.5%) presented with cardiac symptoms, such as chest tightness, palpitations, dizziness, syncope, fatigue, and decreased endurance.Three cases(1.4%) had atypical symptoms of paroxysmal crying, dyspnoea and cyanosis, and 48 cases(23.1%) were asymptomatic.Levels of serological markers of myocardial injury were elevated in 59 cases(28.4%), and 140 cases(67.3%) had predominant ST-T abnormalities on electrocardiograms.Transthoracic echocardiography identified 27 cases (13%) with cardiac enlargement and ten cases(4.8%) with left ventricular systolic dysfunction.There were 126 cases(60.6%) with the anomalous origin of the left coronary artery revealed by CTCA, 50 cases(24.0%) with the anomalous origin of the right coronary artery and 32 cases(15.4%) with bilateral coronary arteries of anomalous origin.Five children underwent surgical treatment, and the remaining 203 children were treated conservatively with drugs.The whole group was successfully treated, and no death case was recorded during the follow-up period.Conclusion:AOCA may cause different degrees of myocardial ischemia.Diverse clinical presentations and diagnostic limitations of transthoracic echocardiography often lead to missed diagnosis or misdiagnosis.In contrast, CTCA has high diagnostic accuracy and can be used to identify the location and course of the coronary ostia.Hence, the management of AOCA should be tailored on a case-to-case basis, taking into consideration of the specific type of coronary origin, with surgical intervention being warranted if necessary.