A rare strain of actinomycetes, with broad-spectrum antimicrobial activity, was isolated from the soil samples from the farmland in the area of Yaohu lake in Nanchang. The information about the taxonomic identification, such as the morphology, physiological properties, cell components and 16S rRNA gene se-quences, suggested that the rare strain of actinomycetes was identified as Micromonospora carbonacea.

Based on the strain of Micromonospora carbonacea JXNU-1 with board-spectrum antimicrobial activity, the technology for the isolation and purification of antibiotic from the fermentation broth of the Micromonospora carbonacea, and its physical-chemical properties were studied. The results showed that, the antibiotic was stable under the condition of high temperature and alkali, but not in acid solution. After the pretreatment of centrifugation and filtration to remove the cells and lipids, the antibiotic was absorbed to negative exchange resin, and the impurity was excluded when 2 mol/L NaCl was used as primary eluent. The antibiotic could be eluted with 20% alcohol as eluent, and the eluting speed of the antibiotic was greatly accelerated as 2 mol/L NaCl was added into 20% alcohol as final eluent. Aqueous solution of the antibiotic was yielded from the alcohol-salt eluant by decompression concentration to wipe off alcohol and by dialysis to exclude salt. One active component was detected in antibiotic solution by paper chromatography, and theHPLC purity was over 99%. As the antibiotic shows positive color-forming reaction to Molish reagents, Benedict’s reagents and Diohenvlamine reagents, combined with the characteristics of absorption spectra, it is deduced that the antibiotic belongs to nucleoside antibiotics.

Advances in mechanism and application of the heating effect in breeding of microorganism are reviewed in this paper. Heat produces mutagenesis effect and screening effect. Heating mutagenesis effect is occurred through the substitution of G-C base pair induced by heat, and heating screening effect produces higher forward mutation rate induced by other mutagens.

Recently, the immunotherapy has been highlighted among cancer treatments. Cancer-testis antigen (CTA) has been studied in a variety of solid tumors because of its specific expression in tumors, and testis, ovary and placenta tissues, but not in other normal tissues. In order to provide a new approach for multiple myeloma (MM) immunotherapy, we examined the CTA expression in MM cell lines, and primary myeloma cells in patients with MM. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of MAGE-C1/CT7, SSX1, SSX2 and SSX4 in MM cell lines of RPMI-8226 and U266, and bone marrow (BM) cells of 25 MM patients and 18 healthy volunteers. The results showed that the 4 CTAs were expressed in RPMI-8226 and U266 cell lines. The positive expression rate of MAGE-C1/CT7, SSX1, SSX2 and SSX4 in the BM cells of 25 MM patients was 28% (7/25), 80% (20/25), 40% (10/25) and 68% (17/25), respectively. In contrast, the expression of any member of the CTAs was not detected in BM cells of 18 healthy volunteers. The expression of two or more CTAs was detected in 80% (20/25) MM patients, and that of at least one CTA in 88% (22/25). The mRNA expression levels of SSX1 and SSX4 were significantly higher in patients with MM at stage III than in those at stage I and II (P<0.05). No statistically significant differences were observed in the mRNA expression levels of MAGE-C1/CT7 and SSX2 in further stratified analyses by age, gender, MM types and percentage of MM cells in BM (P>0.05). In conclusion, our present study showed that MAGE-C1/CT7, SSX1, SSX2 and SSX4 were co-expressed in MM cell lines and the primary myeloma cells in MM patients, but not expressed in BM cells of healthy subjects. The mRNA levels of SSX1 and SSX4 are associated with MM clinical stage. This work may provide a new insight into MM immunotherapy in the future.

BACKGROUNDThe study was designed to investigate the status of molecular epidemiology of HCMV in Urumqi through genetic comparison of clinical isolates.

METHODSDNA sequences of 2.0-2.6 kb were amplified by polymerase chain reaction from three relatively conservative gene regions (DNA polymerase, glycoproteins H, and major immediate-early antigen) of 28 clinical HCMV strains and then were analysed by restriction enzymes.

RESULTSThe restriction patterns of the clinical isolates which did not have relation in epidemiology were greatly different, but the patterns of the clinical isolates related in epidemiology such as strains paired in mother and infant were quite similar. Of eight mother and infant pairs, from whom HCMV were isolated, four pairs showed identity of restriction profiles within each pair for all three amplified regions, four pairs showed differences between mother and infant.

CONCLUSIONThese results confirm the high degree of genetic variability among cytomegalovirus strains in Urumqi. Analysis of PCR-RFLP can indicate transmission of HCMV infection and facilitate its molecular epidemiologic studies.

China ; epidemiology ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; epidemiology ; virology ; DNA-Directed DNA Polymerase ; genetics ; Humans ; Immediate-Early Proteins ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Viral Envelope Proteins ; genetics

In this paper, the five strains of Polygonatum odoratum were used as the experimental materials to test the supercooling point, freezing point, the degree of supercooling, the transition stage time, cooling time and water composition of the plant tissue. The cold resistance of P. odoratum was analyzed with the Gray Correlation Method. The results showed that the cold resistances of the five strains of P. odoratum were different, and the water content of plant tissue had some relevance with freezing point and supercooling point, whereas, it could not be measured when the moisture content was too low. The order of cold resistance of the five strains of P. odoratum was ZJCY, DYYZ, XYYZ, CYYZ and JZ I.
Cold Temperature ; Plant Roots ; chemistry ; physiology ; Polygonatum ; chemistry ; classification ; physiology ; Water ; analysis

OBJECTIVE: To study the protective effect of asiaticoside against hyperoxia-induced bronchopulmonary dysplasia in neonatal rats based on the microRNA-155 (miR-155)/suppressor of cytokine signaling-1 (SOCS1) axis. METHODS: Neonatal rats were randomly divided into a control group, a model group, a low-dose asiaticoside group (10 mg/kg), a middle-dose asiaticoside group (25 mg/kg), a high-dose asiaticoside group (50 mg/kg), and a budesonide group (1.5 mg/kg), with 12 rats in each group. All rats except those in the control group were exposed to a high concentration of oxygen for 14 days to establish a neonatal rat model of bronchopulmonary dysplasia. The low-, middle-, and high-dose asiaticoside groups were given asiaticoside at different doses by gavage, and those in the budesonide group were given budesonide aerosol treatment. Hematoxylin and eosin staining was used to observe lung tissue development and measure radial alveolar count (RAC) and mean linear intercept (MLI). Superoxide dismutase (SOD) and malondialdehyde (MDA) detection kits were used to measure the levels of SOD and MDA in lung tissue. ELISA was used to measure the serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Quantitative real-time PCR was used to measure the mRNA expression of miR-155 and SOCS1 in lung tissue. Western blotting was used to measure the protein expression of SOCS1 in lung tissue. RESULTS: Compared with the control group, the model group had the symptoms of bronchopulmonary dysplasia such as a disordered structure of lung tissue, enlargement of alveolar fusion, uneven alveolar septa, enlargement of average alveolar space, and a reduction in alveolar number. The model group also had significant increases in MLI, MDA level in lung tissue, serum levels of IL-6 and TNF-α, and miR-155 level in lung tissue (P<0.05) and significant reductions in RAC, SOD level, and mRNA and protein expression of SOCS1 in lung tissue (P<0.05). Compared with the model group, the low-, middle-, and high-dose asiaticoside groups and the budesonide group had significant improvement in the above symptoms of bronchopulmonary dysplasia, significant reductions in MLI, MDA level in lung tissue, serum levels of IL-6 and TNF-α, and miR-155 level in lung tissue (P<0.05), and significant increases in RAC, SOD level, and mRNA and protein expression of SOCS1 in lung tissue (P<0.05). Asiaticoside improved the above symptoms and indices in a dose-dependent manner. There were no significant differences in the above indices between the high-dose asiaticoside and budesonide groups (P>0.05). CONCLUSIONS Asiaticoside can alleviate inflammation injury induced by hyperoxia in neonatal rats and improve the symptoms of bronchopulmonary dysplasia in a dose-dependent manner, possibly by down-regulating the expression of miR-155 and up-regulating the expression of SOCS1.
Animals ; Animals, Newborn ; Bronchopulmonary Dysplasia ; Hyperoxia ; Lung ; MicroRNAs ; Rats ; Triterpenes

OBJECTIVE: To study the expression and diagnostic value of plasma miR-145 and miR-183 in children with lupus nephritis (LN). METHODS: A total of 92 children with LN who were admitted from January 2016 to May 2019 were enrolled as the LN group, among whom 17 had type II LN, 15 had type III LN, 36 had type IV LN, 18 had type V LN, and 6 had type VI LN. Forty healthy children who underwent physical examination were enrolled as the healthy control group. According to Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), the 92 children with LN were further divided into a stable LN group with 34 children (SLEDAI score <10) and an active LN group with 58 children (SLEDAI score ≥10). RT-PCR was used to measure the expression of miR-145 and miR-183 in plasma. The receiver operating characteristic (ROC) curve was used to analyze the value of plasma miR-145, miR-183, and anti-dsDNA antibody in the diagnosis of LN. Pearson correlation analysis was used to investigate the correlation of the expression levels of miR-145 and miR-183 in plasma with laboratory markers. RESULTS: The LN, active LN, and stable LN groups had significantly higher levels of anti-dsDNA antibody, C-reactive protein, serum creatinine (Scr), and blood urea nitrogen (BUN) than the control group (P<0.05). The active LN group had significantly higher SLEDAI score, anti-dsDNA antibody, Scr, and BUN than the stable LN group (P<0.05). The LN, active LN, and stable LN groups had significantly lower levels of complement C3, complement C4, and serum albumin (Alb) than the control group (P<0.05). The active LN group had a significantly lower level of Alb than the stable LN group (P<0.05). The LN, active LN, and stable LN groups had significantly lower plasma levels of miR-145 and miR-183 than the control group (P<0.01). The active LN group had significantly lower plasma levels of miR-145 and miR-183 than the stable LN group (P<0.01). The children with difference types of LN had significantly lower plasma levels of miR-145 and miR-183 than the control group (P<0.01), and the type V-VI group and the type IV group had significantly lower plasma levels of miR-145 and miR-183 than the type II-III group (P<0.01). The ROC curve analysis showed that the optimal cut-off values of plasma miR-145, miR-183, and anti-dsDNA antibody were 1.05, 0.62, and 186.30 IU/mL respectively, in the diagnosis of LN, and the combination of these three indices had the largest area under the ROC curve of 0.896 (95%CI: 0.835-0.955), with a sensitivity of 90.5% and a specificity of 84.2%. In the children with LN, the plasma levels of miR-145 and miR-183 were negatively correlated with SLEDAI score, anti-dsDNA antibody, Scr, and BUN (P<0.05) and were positively correlated with complement C3, complement C4, and Alb (P<0.05). CONCLUSIONS There are significant reductions in the expression levels of miR-145 and miR-183 in plasma in children with LN, which are correlated with the activity level and pathological typing of LN. Combined measurement of miR-145, miR-183, and anti-dsDNA antibody has a high value in the diagnosis of LN.
Biomarkers ; Child ; Complement C4 ; Humans ; Lupus Nephritis ; genetics ; MicroRNAs ; genetics ; ROC Curve

AIM:To observe the effect of Tangshenfang(TS)on the liver protection and the levels of silent in-formation regulator 1(SIRT1)and peroxisom proliferator-activated receptor γcoactivator-1α(PGC-1α)in the liver tissue. METHODS:The rat model of diabetes mellitus(DM)was established by intravenous injection of streptozotocin(STZ;30 mg/kg)after having the high fat/high glucose diets for 1 month.The diabetic rats were randomly divided into DM group,DM with high-dose TS(TSHi)group, medium-dose TS(TSMed)group and low-dose TS(TSLow)group.The normal rats were served as control group.There were 8 rats in each group.After treatment with TS for 12 weeks,the serum biochemical indi-ces including fasting blood glucose(FBG), triglyceride(TG), alanine aminotransferase(ALT)and aspartate aminotrans-ferase(AST)were tested.Fasting insulin(FINS)was also detected by radioimmunoassay,and homeostatic model assess-ment for insulin resistance(HOMA-IR)was calculated.The serum levels of tumor necrosis factor-α(TNF-α)and interleu-kin-1(IL-1)were measured by ELISA.The activity of SOD and content of MDA in the liver tissues were measured by the methods of hydroxylamine and thiobarbituric acid.The liver pathological changes were observed under light microscope with HE and Masson staining.The protein expression of SIRT1and PGC-1αin the liver tissues was determined by Western blot. RESULTS:In DM group,serum FBG,TG,ALT,AST,FINS,HOMA-IR,TNF-αand IL-1 were obviously increased com-pared with the control group(P<0.01).The fatty changes,local necrosis,inflammation and fibrosis in the liver tissues were observed.The content of MDA in liver increased,while the activity of SOD decreased markedly.The protein expression of SIRT1 and PGC-1αwas decreased(P<0.05).In TS treatment groups,all these changes in DM rats were markedly reversed by TS,and the protein expression of SIRT1 and PGC-1αin the liver tissues was markedly increased.CONCLUSION:TS may protect the rats from diabetic liver injury by increasing the expression of SIRT 1 and PGC-1α,and thereby improving in-sulin resistance and oxidative stress.

OBJECTIVETo explore the optimum conditions for preparing poly(lactic-co-glycolic) acid (PLGA) nanoparticles and evaluate the bioactivity of hepatocyte growth factor (HGF)-loaded PLGA nanoparticles.

METHODSBovine serum albumin (BSA)-loaded PLGA nanoparticles were prepared using a double emulsion-solvent evaporation method. The preparation process of nanoparticles was optimized by orthogonal test with the particle size, encapsulation efficiency (EE), drug loading (DD), and recovery as the indexes. HGF-loaded nanoparticles were then prepared under the optimized conditions. The EE, DD and release characteristics of BSA?loaded nanoparticles and HGF-loaded nanoparticles were evaluated using a BCA kit and HGF ELISA kit. The bioactivity of HGF-loaded nanoparticles was evaluated using CCK8 proliferation assay.

RESULTSThe HGF-loaded nanoparticles prepared under the optimized conditions had a uniform size with a mean diameter of 234.4∓4.8 nm, an EE of (77.75∓3.04)% and a recovery rate of (49.33∓9.34)%. The in vitro release curve highlighted an initial burst drug release followed by sustained release from the nanoparticles. HGF-loaded nanoparticles obviously promoted the proliferation of Hacat keratinocytes in vitro.

CONCLUSIONHGF-loaded nanoparticles prepared using double emulsion?solvent evaporation method under optimized conditions possesses a high EE with a good sustained drug release profile and a good bioactivity.