Effectiveness of tofacitinib combined with iguratimod in the treatment of difficult-to-treat moderate-to-severe rheumatoid arthritis / 北京大学学报(医学版)

OBJECTIVE@#To investigate the efficacy and safety of iguratimod combined with tofacitinib in patients with difficult-to-treat moderate-to-severe rheumatoid arthritis (RA).@*METHODS@#In this prospective clinical study, 30 patients with difficult-to-treat moderate-to-severe RA who attended the Department of Rheumatology and Immunology of Shanxi Province Fenyang Hospital from September 2021 to June 2022 were selected. Twenty-three patients enrollment had been treated with 2 or more conventional synthetic disease modifying anti-rheumatic drugs (DMARDs) for more than 6 months. At least, methotrexate or leflunomide was included. Seven patients were treated with conventional synthetic DMARDs combined with tumor necrosis factor antagonists. Because all the patients had not reached the target of treatment, the combination treatment regimen of DMARDs was changed to iguratimod and tofacitinib. The observation period was 12 weeks. Clinical data were collected before and after treatment. At the end of 4 weeks, 8 weeks and 12 weeks, the clinical data were collected such as swollen joints count (SJC), tender joints count (TJC), time of morning stiffness, clinical disease activity index (CDAI), health status assessment questionnaire (HAQ), and 28-joint disease activity score (DAS28) were included. We collected laboratory indicators, recorded the patient's medication, and observed some changes to see if any adverse drug reactions occurred during the treatment.@*RESULTS@#There were significant differences in erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), platelet (PLT), SJC, TJC, DAS28 based on ESR(DAS28-ESR), time of morning stiffness, HAQ, CDAI, and anti-cyclic citrullinated peptide antibody before and after treatment. The differences had statistical significance (P < 0.05). There was no statistical differences in globulin before and after treatment (P>0.05). During the treatment of iguratimod combined with tofacitinib, there was no serious adverse reactions such as leukopenia, significant elevation of liver enzymes, allergy or thromboemblolic events that occurred in all the patients.@*CONCLUSION@#Iguratimod combined with tofacitinib in the treatment of difficult-to-treat moderate-to-severe RA may have efficacy. The machanism was improving the patients' recent clinical symptoms by reducing inflammatory indexes. This combination treatment regimen with iguratimod and tofacitinib has a good safety profile.

Up-regulation of integrin α6ß4 expression by mitogens involved in dairy cow mammary development.

In dairy cows, the extracellular microenvironment varies significantly from the virgin state to lactation. The function of integrin α6ß4 is dependent on cell type and extracellular microenvironment, and the precise expression profile of α6ß4 and its effects on mammary development remain to be determined. In the present study, real-time PCR and immunohistochemistry were used to analyze the expression and localization of integrin α6ß4 in Holstein dairy cow mammary glands. The effects of integrin α6ß4 on the proliferation induced by mammogenic mitogens were identified by blocking integrin function in purified dairy cow mammary epithelial cells (DCMECs). The results showed that the localization of ß4 subunit and its exclusive partner the α6 subunit were not consistent but were co-localized in basal luminal cells and myoepithelial cells, appearing to prefer the basal surface of the plasma membrane. Moreover, α6 and ß4 subunit messenger RNA (mRNA) levels changed throughout the stages of dairy cow mammary development, reflected well by protein levels, and remained higher in the virgin and pregnancy states, with duct/alveolus morphogenesis and active cell proliferation, than during lactation, when growth arrest is essential for mammary epithelial cell differentiation. Finally, the upregulation of integrin expression by both mammogenic growth hormone and insulin-like growth factor-1 and the inhibited growth of DCMECs by function-blocking integrin antibodies confirmed that integrin α6ß4 was indeed involved in dairy cow mammary development.

Research progress of trans-cinnamaldehyde pharmacological effects / 中国中药杂志

Trans-cinnamaldehyde, the main component of volatile oil from cassia twig or Cinnamomum cassia, which is a traditional Chinese herbal medicine. Trans-cinnamaldehyde is a kind olefine aldehyde of organic compounds and has many pharmacological properties, such as anti-inflammatory, anti-tumor, anti-bacterial, antidiabetic, anti-obesity, and neuroprotection etc. The compound has preventive and therapeutic effects on the nervous system, cardiovascular, cancer, diabetes and other diseases. Trans-cinnamaldehyde, as a preventive care of nature medicine, has great clinical and market potential. This paper gives a review about the pharmacological effects and mechanism of trans-cinnamaldehyde researched in the latest five years. We hope to provide some basic information for further research on trans-cinnamaldehyde.

Function of SREBP1 in the milk fat synthesis of dairy cow mammary epithelial cells.

Sterol regulatory element-binding proteins (SREBPs) belong to a family of nuclear transcription factors. The question of which is the most important positive regulator in milk fat synthesis in dairy cow mammary epithelial cells (DCMECs) between SREBPs or other nuclear transcription factors, such as peroxisome proliferator-activated receptor γ (PPARγ), remains a controversial one. Recent studies have found that mTORC1 (the mammalian target of rapamycin C1) regulates SREBP1 to promote fat synthesis. Thus far, however, the interaction between the SREBP1 and mTOR (the mammalian target of rapamycin) pathways in the regulation of milk fat synthesis remains poorly understood. This study aimed to identify the function of SREBP1 in milk fat synthesis and to characterize the relationship between SREBP1 and mTOR in DCMECs. The effects of SREBP1 overexpression and gene silencing on milk fat synthesis and the effects of stearic acid and serum on SREBP1 expression in the upregulation of milk fat synthesis were investigated in DCMECs using immunostaining, Western blotting, real-time quantitative PCR, lipid droplet staining, and detection kits for triglyceride content. SREBP1 was found to be a positive regulator of milk fat synthesis and was shown to be regulated by stearic acid and serum. These findings indicate that SREBP1 is the key positive regulator in milk fat synthesis.

Functional analysis of FABP3 in the milk fat synthesis signaling pathway of dairy cow mammary epithelial cells.

Milk fat is the major energy component of milk, and regulation of its production relies on transcription factors sterol regulatory element-binding protein 1 (SREBP1) and peroxisome proliferator-activated receptor gamma (PPARγ). As one of the target genes of SREBP1 and PPARγ, fatty acid-binding protein 3(FABP3) is the main protein allowing for rapid diffusion and selective targeting of long-chain fatty acids toward specific organelles for metabolism. Whether FABP3 plays an important role in milk fat synthesis signaling pathway is largely unknown. In this study, we observed the effects of FABP3 overexpression and gene silencing in dairy cow mammary epithelial cells, as well as the effects of oleic acid, stearic acid, and palmitic acid on the expressions of FABP3 and lipid droplet formation, by using quantitative reverse transcriptase (qRT)-PCR, Western blotting, and fluorescent immunostaining techniques. FABP3 upregulated the expression of SREBP1 and PPARγ to increase lipid droplet accumulation. Oleic acid, stearic acid, and palmitic acid also increased lipid droplet accumulation by affecting expression of FABP3. These findings shed new insights for understanding the mechanism of FABP3 in regulating milk fat synthesis.

Enhanced effect of guizhi plus Gegen Decoction on learning and memory disorder in LPS induced neuroinflammatory mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the potential effect of Guizhi plus Gegen Decoction (GGD) in improving learning and memory of lipopolysaccharides (LPS) induced neuroinflammatory mice and its possible mechanisms.</p><p><b>METHODS</b>Totally 63 male ICR mice were randomly divided into 5 groups, i.e., the normal control (n = 13), the model group (n = 13), the low dose GGD group (n = 10), the high dose GGD group (n = 14), and the positive control group (n = 13). Mice were intraperitoneally injected with LPS (0.33 mg/kg) to induce Alzheimer's disease (AD) model. Mice in the high and the low dose GGD groups were administered with 12 g/kg or 6 g/kg by gastrogavage for 4 successive weeks. Mice in the control group were intraperitoneally injected with minocycline (50 mg/kg) for 3 days. By the end of treatment LPS were injected 4 h before behavior test each day, and then behavior test was conducted in mice of each group. Effect of GGD on learning and memory of AD mice was observed by using open field test, novel object recognition task, and Morris water maze.</p><p><b>RESULTS</b>Open field test showed there was no statistical difference in the movement time and the movement distance among all groups (P > 0.05), suggesting that LPS and GGD had no effect on locomotor activities of mice. In novel object recognition test, AD mice spent significantly shorter time to explore novel object after they were induced by LPS (P < 0.05), while for AD mice in the low and high dose GGD groups, their capacities for exploration and memory were significantly improved (P < 0. 05, P < 0.01). Results of Morris water maze showed that AD mice exhibited increased escape latency (P < 0.05) and spent much less time in swimming across the original platform (both P < 0.05). However, AD mice in the low and high dose GGD groups had obvious shortened latency and increased time percentage for swimming (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>GGD possessed certain improvement in learning and memory disorder of LPS induced AD mice.</p>

SOCS3-mediated blockade reveals major contribution of JAK2/STAT5 signaling pathway to lactation and proliferation of dairy cow mammary epithelial cells in vitro.

Suppressor of cytokine signaling 3 (SOCS3) is a cytokine-induced negative feedback-loop regulator of cytokine signaling. More and more evidence has proved it to be an inhibitor of signal transducers and activators of transcription 5 (STAT5). Here, we used dairy cow mammary epithelial cells (DCMECs) to analyze the function of SOCS3 and the interaction between SOCS3 and STAT5a. The expression of SOCS3 was found in cytoplasm and nucleus of DCMECs by fluorescent immunostaining. Overexpression and inhibition of SOCS3 brought a remarkable milk protein synthesis change through the regulation of JAK2/STAT5a pathway activity, and SOCS3 expression also decreased SREBP-1c expression and fatty acid synthesis. Inhibited STAT5a activation correlated with reduced SOCS3 expression, which indicated that SOCS3 gene might be one of the targets of STAT5a activation, DCMECs treated with L-methionine (Met) resulted in a decrease of SOCS3 expression. SOCS3 could also decrease cell proliferation and viability by CASY-TT detection. Together, our findings indicate that SOCS3 acts as an inhibitor of JAK2/STAT5a pathway and disturbs fatty acid synthesis by decreasing SREBP-1c expression, which validates its involvement in both milk protein synthesis and fat synthesis. In aggregate, these results reveal that low SOCS3 expression is required for milk synthesis and proliferation of DCMECs in vitro.

Goat mammary gland expression of Cecropin B to inhibit bacterial pathogens causing mastitis.

The antibacterial peptide Cecropin B (CB), isolated from the giant silk moth, has been shown to effectively eliminate bacteria. In this study, the effects of transgenic CB on dairy goat mammary epithelial cells (DGMECs) and dairy goat mammary gland were investigated. The DNA of CB from silkworm was amplified by reverse transcription PCR (RT-PCR) and then fused to the eukaryotic expression vector pECFP-C1. The recombinant plasmid pECFP-Cecropin B (pECFP-CB) was used for the transfection of DGMECs, and the expression of transgenic CB and the antibacterial activity of it were confirmed by western blot and agar diffusion reaction respectively. The stable DGMEC line transfected by pECFP-CB was obtained by screening with G418. In vivo experiment, pECFP-CB was injected into dairy goat mammary gland, and also the expression and antibacterial activity of transgenic CB were confirmed. Results of this study: transgenic CB can be expressed in DGMECs and dairy goat mammary gland, and inhibit the mastitis caused by Staphylococcus aureus.

Proteomic and functional analyses reveal MAPK1 regulates milk protein synthesis.

L-Lysine (L-Lys) is an essential amino acid that plays fundamental roles in protein synthesis. Many nuclear phosphorylated proteins such as Stat5 and mTOR regulate milk protein synthesis. However, the details of milk protein synthesis control at the transcript and translational levels are not well known. In this current study, a two-dimensional gel electrophoresis (2-DE)/MS-based proteomic technology was used to identify phosphoproteins responsible for milk protein synthesis in dairy cow mammary epithelial cells (DCMECs). The effect of L-Lys on DCMECs was analyzed by CASY technology and reversed phase high performance liquid chromatography (RP-HPLC). The results showed that cell proliferation ability and ß-casein expression were enhanced in DCMECs treated with L-Lys. By phosphoproteomics analysis, six proteins, including MAPK1, were identified up-expressed in DCMECs treated with 1.2 mM L-Lys for 24 h, and were verified by quantitative real-time PCR (qRT-PCR) and western blot. Overexpression and siRNA inhibition of MAPK1 experiments showed that MAPK1 upregulated milk protein synthesis through Stat5 and mTOR pathway. These findings that MAPK1 involves in regulation of milk synthesis shed new insights for understanding the mechanisms of milk protein synthesis.

Stat5a increases lactation of dairy cow mammary gland epithelial cells cultured in vitro.

Signal transducer and activator of transcription 5a (Stat5a) transduces signals of extracellular cytokines and growth factors to the nucleus of mammary gland epithelial cells and thereby regulates gene transcription during pregnancy, lactation, and weaning. However, its function on the milk production of dairy cows needs further investigation. In this experiment, the effects of Stat5a on lactation ability of dairy cow mammary gland epithelial cells (DCMECs) were analyzed. Eukaryotic expression vector pcDNA3.1+-stat5a-αS1 was constructed by inserting stat5a gene into the plasmid vector pcDNA3.1+ and replacing CMV promoter with α-S1-casein 5' flanking sequence. The recombinant vector was stably transfected into DCMECs after geneticin (G418) selection. The proliferation and viability of DCMECs, expression of ß-casein and stat5a gene, and the content of lactose were detected. The results showed that stat5a gene in eukaryotic expression vector pcDNA3.1+-stat5a-αS1 was highly expressed in DCMECs and could increase the lactation ability of DCMECs. The associativity of Stat5a with nutrients on the lactation ability of DCMECs was also evaluated. Lysine (Lys), methionine (Met), sodium acetate, ß-sodium hydroxybutyrate, and glucose all had more positive effects on the lactation function of DCMECs after pcDNA3.1+-stat5a-αS1 transfection. The proliferation and viability of DCMECs, expression of ß-casein and stat5a gene, and contents of lactose and triglyceride were detected. The results revealed that nutrients could promote expression of Stat5a gene to increase lactation of DCMECs. These data help to clarify the function of stat5 gene on lactation and gene regulatory networks linking stat5a.

MiR-15a decreases bovine mammary epithelial cell viability and lactation and regulates growth hormone receptor expression.

MicroRNAs (miRNAs) are a class of small non-coding RNAs that regulate the expression of target genes at the post-transcriptional level by transcript degradation or translational inhibition. The role of bta-miR-15a in bovine mammary gland hasn't been reported. Using miRNAs prediction software, GHR gene was predicted to be a potential target of bta-miR-15a. In this study, bovine mammary epithelial cell line was used as an in vitro cell model to address the function of bta-miR-15a on bovine mammary epithelial cells. The expression changes of bta-miR-15a and Ghr after bta-miR-15a transfection were detected by qRT-PCR; the expression of GHR protein and casein was detected by western blotting. To determine whether bta-miR-15a can affect cell viability, cells were examined using an electronic Coulter counter (CASY-TT). In conclusion, bta-miR-15a inhibited the expression of casein of bovine mammary epithelial cells, and cell number and viability were reduced by bta-miR-15a expression. Bta-miR-15a inhibited the viability of mammary epithelial cells as well as the expression of GHR mRNA and protein level, therefore suggesting that bta-miR-15a may play an important role in mammary gland physiology.

Proteomic analysis of the nuclear phosphorylated proteins in dairy cow mammary epithelial cells treated with estrogen.

Estrogen regulates a variety of physiological processes, including mammary gland growth, morphogenesis of the mammary gland, proliferation and differentiation, and elevating the expression of milk proteins. Many nuclear phosphorylated proteins such as pStat5 and mTOR regulate milk protein synthesis. But the detail of milk protein synthesis controlled at the transcript level and posttranslational level is not well-known. To contribute to the understanding of the molecular mechanism underlying estrogen action on the dairy cow mammary epithelial cells (DCMECs), nuclear phosphorylated proteins regulated by estrogen in DCMECs were identified. Two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization/time of flight mass spectrometry were used to identify the changes of nuclear phosphorylated proteins in DCMECs treated with estrogen. Seven proteins were identified differentially up-expressed in DCMECs after 24-h estrogen exposure: including glycyl-tRNA synthetase, previously reported in milk protein synthesis of DCMECs, belonging to the class-II aminoacyl-tRNA synthetase family; proteins involved in other cellular functions, such as translation initiation factors, GTP-binding nuclear proteins, heat-shock proteins, and proteins belonging to ubiquitin-proteasome system. This screening reveals that estrogen influences the levels of nuclear phosphorylated proteins of DCMECs which opens new avenue for the study of the molecular mechanism linking to milk synthesis.

Establishment and characterization of a lactating dairy goat mammary gland epithelial cell line.

To study milk synthesis in dairy goat mammary gland, we had established an in vitro lactating dairy goat mammary epithelial cell (DGMEC) line. Mammary tissues of Guan Zhong dairy goats at 35 d of lactation were dispersed and cultured in a medium containing epithelial growth factor, insulin-like growth factor-1, insulin transferrin serum, and fetal bovine serum. Epithelial cells were enriched by digesting with 0.25% trypsin repeatedly to remove fibroblast cells and were identified as epithelial origin by staining with antibody against cytokeratine 18. The DGMECs displayed monolayer, cobble-stone, epithelial-like morphology, and formed alveoli-like structures and island monolayer aggregates which were the typical characteristics of mammary epithelial cells. A one-half logarithmically growth curve and cytoplasmic lipid droplets in these cells were observed. In this paper, we also studied the lactating function of DGMECs. Results showed that DGMECs could secrete lactose and ß-casein. Lactating function of the cells had no obvious change after 48 h treated by insulin, while prolactin could obviously raise the secretion of milk proteins and lactose.

Changes of endoplasmic reticulum and mitochondria in mammary epithelial cells during mammogenesis in Chinese Holstein dairy cows.

The objective of the research was to study the changes of the major organelles, endoplasmic reticulum (ER) and mitochondria, in mammary epithelial cells of the Chinese Holstein dairy cow during mammogenesis. For this purpose, a mammary epithelial cell bank was established from 9 selected Chinese Holstein dairy cows using collagenase I digestion and attachment culture biotechniques. This cell bank included 9 samples at stages of pregnancy, lactation and involution. The changes of ER and mitochondria in the mammary cells were observed at the subcellular level using living cell fluorescent labeling and laser confocal microscopy. Subsequently, the area of integrated optical density of each sample was calculated to determine changes of ER and mitochondria in the mammary epithelial cells. The results showed clear differences in the epithelial major organelles during the various mammary gland development stages. The ER and mitochondria, as an indicator of lactogenic activity of alveolar secretory cells, increased in number from pregnancy to lactation by an average 37.32% and 18.44%, respectively, which was followed by a reduction at involution by an average 38.04% and 22.91% compared to lactation. Our study shows that the stages of mammogenesis are accompanied by changes in activity of the major organelles of the mammary epithelial cells.

Fragile X syndrome and epilepsy.

Fragile X syndrome (FXS) is one of the most prevalent mental retardations. It is mainly caused by the loss of fragile X mental retardation protein (FMRP). FMRP is an RNA binding protein and can regulate the translation of its binding RNA, thus regulate several signaling pathways. Many FXS patients show high susceptibility to epilepsy. Epilepsy is a chronic neurological disorder which is characterized by the recurrent appearance of spontaneous seizures due to neuronal hyperactivity in the brain. Both the abnormal activation of several signaling pathway and morphological abnormality that are caused by the loss of FMRP can lead to a high susceptibility to epilepsy. Combining with the research progresses on both FXS and epilepsy, we outlined the possible mechanisms of high susceptibility to epilepsy in FXS and tried to give a prospect on the future research on the mechanism of epilepsy that happened in other mental retardations.

Translocation of phospho-protein kinase Cs implies their roles in meiotic-spindle organization, polar-body emission and nuclear activity in mouse eggs.

The protein kinase Cs (PKCs) are a family of Ser/Thr protein kinases categorized into three subfamilies: classical, novel, and atypical. The phosphorylation of PKC in germ cells is not well defined. In this study, we described the subcellular localization of phopho-PKC in the process of mouse oocyte maturation, fertilization, and early embryonic mitosis. Confocal microscopy revealed that phospho-PKC (pan) was distributed abundantly in the nucleus at the germinal vesicle stage. After germinal vesicle breakdown, phospho-PKC was localized in the vicinity of the condensed chromosomes, distributed in the whole meiotic spindle, and concentrated at the spindle poles. After metaphase I, phospho-PKC was translocated gradually to the spindle mid-zone during emission of the first polar body. After sperm penetration and electrical activation, the distribution of phospho-PKC was moved from the spindle poles to the spindle mid-zone. After the extrusion of the second polar body (PB2) phospho-PKC was localized in the area between the oocyte and the PB2. In fertilized eggs, phospho-PKC was concentrated in the pronuclei except for the nucleolus. Phospho-PKC was dispersed after pronuclear envelope breakdown, but distributed on the entire spindle at mitotic metaphase. The results suggest that PKC activation may play important roles in regulating spindle organization and stabilization, polar-body extrusion, and nuclear activity during mouse oocyte meiosis, fertilization, and early embryonic mitosis.

Morphological changes to early stage Taenia solium cysticerci following oxfendazole treatment.

The progressive morphological changes to early stage Taenia solium cysticerci following the treatment of pigs with a single therapeutic dose of oxfendazole (30 mg/kg), are described. On Day 1 after treatment, no obvious changes occurred in the general appearance of the larvae but alternations were seen by electron microscope, with an apparent reduction in the number of microtriches, and a complete disappearance of the tegument. Numerous granules were seen to have accumulated in the tegument cells. As treatment progressed, damage to the cysticerci was more serious and, by five days, all cysticerci were seen to be in an advanced stage of degeneration. By 45 days post-treatment, all cysts were calcified. These results suggest that oxfendazole is a highly effective drug against T. solium cysticerci in the early stages of development.

Evaluation for clinical staging parameters of prostate neoplasm / 中华男科学杂志

<p><b>OBJECTIVES</b>To evaluate significance of clinical parameters in prostate cancer staging.</p><p><b>METHODS</b>One hundred and twelve patients of prostate cancer were diagnosed by transrectal ultrasound-guided prostate needle biopsies. These cases were staged by pathologic diagnosis, MRI and bone scan. Clinical significance of serum PSA, Gleason score of biopsy, percentage of positive biopsy cores in prostate cancer staging were evaluated.</p><p><b>RESULTS</b>Of 112 patients, 30.4% (34/112) underwent radical retropubic prostatectomy. The serum PSA, Gleason score of biopsy and percentage of positive biopsy cores, were significant correlation with staging prostate cancer (r = 0.698, r = 0.674, r = 0.671, P < 0.001), and no significant difference between staging B and staging C (chi 2 = 2.675, P = 0.096; chi 2 = 0.704, P = 0.401). PSA in patients with stage D had significant difference with others (chi 2 = 5.135, P = 0.023; chi 2 = 4.593, P = 0.032). The sensitivity, specificity and accuracy of PSA were 76.7%, 50.0% and 71.4% respectively. Those of Gleason score and percentage of positive biopsy cores were 83.3%, 77.3%, 82.1% and 77.8%, 54.5%, 73.2% respectively.</p><p><b>CONCLUSIONS</b>The serum PSA, Gleason score of biopsy and percentage of positive biopsy cores had clinical significance in the staging of prostate cancer. Gleason score of biopsy in staging was more accurate than that of the other two parameters and the serum PSA can better predict prostate cancer metastasis.</p>

Screening of Flocculant-Producing Bacteria and Application in Treatment of Azo Dye Wastewater / 环境与健康杂志

Objective To screen the flocculatnt-producing strain from activated sludge that can treat azo dyeing wastewater effectively.Methods Screening and rescreening were used to acquire the strain which possesses high efficiency of flocculatant production and the strain was identified.The character of microbial flocculant(MBF) and the ability to treat azo dye wastewater were studied.Results A strain of high-efficiency bioflocculant-producting bacteria(AJ-6),initially identified as Alcaligenes sp,was screened.The bioflocculant produced by the strain was able to flocculate kaolin suspension with 94.4% and fly ash suspension with 98.9% respectively.The flocculation activity distribution tests showed that the active components of the bioflocculant existed in the supernatant fluid after centrifugation.It had good treatment efficiency in treating azo dyes methyl orange wastewater.The maximal efficiencies of removing CODCr and chroma from the wastewater were 81.3% and 94.2%.Compared with the other flocculants,the performance of MBF was better than that of polyacrylamide(PAM),aluminum sulfate,ferrous sulfate.The MBF was more thermostable when treated with 100 ℃ for 30 min,and the animal toxicity test showed that the MBF had no acute toxicity to mice.Conclusion The bioflocculant produced by the strain AJ-6 is applicable to treat azo dyeing wastewater and can be also used for the other wastewater.