Objective: To screen and identify the dominant strains which produce fibrinolytic enzyme during the processing of Sojae Semen Praeparatum (SSP, Dandouchi in Chinese). Methods: SSP was prepared according to the Chinese Pharmacopoeia (2020 edition), and samples were taken at different time points during the fermenting process of SSP.The casein plate method and fibrin plate method were used to screen the fibrinolytic enzyme-producing microorganisms in samples at different time points. The fibrinolytic enzyme-producing microorganisms were inoculated in the designated liquid medium to obtain single strain fermentation broth, and fibrin plate method was used to measure the fibrinolytic activity of the fermentation broth. The DNA sequences of fibrinolytic enzyme-producing bacteria and fungi were amplified using 16S rDNA and 18S rDNA universal primer by PCR respectively.The amplified products were sequenced, and the sequencing results were identified through NCBI homology comparison. Molecular biological identification was done by phylogenetic tree constructed by MEGA 4.1 software. Results: Three types of fibrinolytic enzyme-producing bacteria were screened out and identified in this study. They were Bacillus subtilis, Stenotrophomonas maltophilia and Micrococcus, respectively. The result of fibrin plate method showed that the fermentation broth of S. maltophilia had the highest fibrinolytic activity, reaching 527.49 IU/mL. Conclusion: There are fibrinolytic enzyme-producing dominant microorganisms existing in the fermenting process of SSP and the thrombolytic effect of SSP is worthy of further study. This study lays the foundation for revealing the formation mechanism of fibrinolytic enzyme in the fermentation process of SSP.

Abstract: Objective To understand the mental health literacy level of residents in Hainan Province, and to provide evidence for promoting mental health promotion and improving the mental health literacy level of residents. Methods Multi-stage random sampling method was used to investigate The National Mental Health Literacy Questionnaire among 6 895 residents in 12 districts including Changjiang, Chengmai, Dongfang, Ledong, Lingshui, Haikou Longhua District, Haikou Meilan District, Qionghai, Sanya, Tunchang, Wenchang and Wuzhishan. Results　In the survey 6 895 residents in 12 counties, cities, districts of Hainan Province, 365 of them reached the standard of mental health with the standard rate of 5.3%. The mental health knowledge score was (54.00±17.02) with the standard rate of 7.7%; the self-assessment score was (26.11±3.99) with the standard rate of 75.9%; the score of mental health skill was (26.22±7.25) with the standard rate of 44.8%. The mental health literacy level of medical workers was the highest, with the standard rate of 13.5% (112/830), and the mental health literacy level of farmers was the lowest, with the standard rate of 0.8% (13/1 647). The results of unconditional Logistic regression showed that the factors entering the regression model included educational background (OR=2.268), personal monthly income (OR=1.129), gender (OR=1.302), household registration (OR=0.776), and whether they had participated in mental health related courses OR training (OR=0.511). The higher the educational background and personal monthly income, the higher the psychological quality. The mental health of women was higher than that of men, and that of urban was higher than that of rural, and those who had participated in mental health related courses were higher than those who had not. Conclusions The mental health literacy level of Hainan residents is at a low level, and the influencing factors are multifaceted. It is suggested to strengthen the mental health monitoring comprehensively and carry out rich health education service mode for different groups.

OBJECTIVEIdiopathic nephrotic syndrome (INS) is a common glomerular disease. The pathogenesis of the disease remains unclear. Recent studies indicate that transforming growth factor beta (TGF beta) is the main cytokine involved in glomerular disease. It plays an important role in the development of INS and in occurrence of glomerulosclerosis. The present study aimed to study changes and significance of TGF beta in children with idiopathic nephrotic syndrome (INS).

METHODSTotally 35 cases with INS (13 males, 22 females) were studied. The age of onset was between 2 years and 1 months and 14 years with an average of 8 years and 3 months. The active stage group had 35 cases and the remission stage groups had 25 cases. The cases in active stage group had first onset of the disease with obvious clinical symptoms and abnormal laboratory findings without use of corticosteroids. The cases in remission stage group were asymptomatic without abnormal laboratory findings. Protein in urine was negative over 4 weeks after oral administration of prednisone for 8 weeks. Twenty five cases were steroid responsive and 10 cases were steroid non-responsive among the 35 cases. Thirty healthy young children were enrolled as control. TGF beta was detected by ELISA in peripheral blood mononuclear cell (PBMC) culture medium. The TGF beta mRNA gene expression was measured by in situ PCR in PBMC.

RESULTS(1) Concentration of TGF beta(247 +/- 26) ng/L and TGF beta mRNA expression (0.57 +/- 0.18) in active stage of simple type or nephritis type INS were higher than those of remission stage and control (P < 0.01). Concentration of TGF beta[(125 +/- 16) ng/L] and TGF beta mRNA expression (0.30 +/- 0.12) in remission stage were higher than that of control (P < 0.05). (2) The level of TGF beta protein in nephritis type [(275 +/- 26) ng/L] was significantly higher than that in simple type [(220 +/- 18) ng/L] in active stage INS (t = 6.45, P < 0.01). No significant difference in TGF beta mRNA expression was found between the nephritis type (0.58 +/- 0.15) and simple type (0.55 +/- 0.16) in active stage INS, either (P > 0.05). But these two types were different from the control (P < 0.01). (3) Concentration of TGF beta and TGF beta mRNA expression after therapy was clearly lower than that before therapy in steroid responsive group (P < 0.01). Whereas no significant change was seen in steroid non-responsive group. Both indicators were higher in steroid non-responsive group than in steroid responsive group whether before or after therapy.

CONCLUSIONTGF beta may play an important role in the mechanism of INS and its level in PBMC can be used as an immunological indicator for the illness state, therefore, determination of TGF beta level and mRNA may be of some clinical significance.

Adolescent ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Nephrotic Syndrome ; blood ; drug therapy ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; genetics ; metabolism

BACKGROUNDGlioma-induced edema is considered as one of the most pathological characteristics of glioma and a significant source of morbidity and mortality. New strategies are needed for the treatment of peritumoral edema in glioma. Endostatin has been proven to be beneficial as an anti-angiogenic agent in experimental gliomas, but the effects are unclear. This study aimed to investigate the effects of endostatin on C6 glioma-induced edema.

METHODSTumorigenic mice were established by subcutaneous injection of three glioma cell lines, C6-null cells and stable transfected-C6 cells overexpressing mock vector (C6-mock cells) and endostatin (C6-endo cells). Endostatin expression in xenograft C6 glioma was determined by immunostaining and Western blotting. Glioma-induced edema and tumor vessel permeability were assayed. The effect of endostatin on vascular enodothelial growth factor (VEGF) expression in vivo was analyzed by quantitative polymerase chain reaction (Q-PCR) and enzyme-linked immunosorbent assay (ELISA). The number of vesiculo-vascuolar organelles (VVOs) formed in tumor endothelia was calculated using electron microscopy. Data were analyzed by using one-way analysis of variance (ANOVA) followed by Dunnett's post hoc test for multiple comparisons to the control groups.

RESULTSOverexpression of endostatin (C6-endo cells) significantly suppressed tumor growth and reduced tumor edema and vessel permeability. ELISA analysis showed that the level of VEGF protein was markedly decreased in tumor from C6-endo cells compared with tumor from C6-null cells and C6-mock cells. Similar results were obtained by Q-PCR. Furthermore, the number of VVOs observed in tumor from C6-endo mice was significantly reduced compared with tumor from C6-null cells or C6-mock cells.

CONCLUSIONSOur data provide primary evidence that endostatin reduces glioma-induced edema and vascular permeability. Using endostatin may be an effective strategy for treating glioma edema.

Animals ; Cell Line, Tumor ; Edema ; drug therapy ; etiology ; Endostatins ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Glioma ; drug therapy ; physiopathology ; Male ; Mice ; Mice, Inbred BALB C ; Polymerase Chain Reaction ; Rats ; Xenograft Model Antitumor Assays

Objective The active ingredient was used as index to optimize the extraction and enrichment process of anti-in-flammatory and analgesic active-fraction(ARF)of Dianbaizhu. Methods Methyl salicylate triglycoside-B was chosen as index com-ponent to extract and enrich methyl salicylate glycosides. Extraction and elution solvents were optimized. The HPLC fingerprint was ob-tained with Thermo Hypersil Gold C18(250 mm×4.6 mm,5μm)column and a gradient elution with the mobile phase consisting of ace-tonitrile(A)-0.2％acetic acid(B)at a flow rate of 1.0 ml/min. And the detection wavelength was set at 294 nm. Results The opti-mized extraction solvent of Dianbaizhu was the 30％ethanol and the optimized elution solvent of ARF enriched by AB-8 macroporous resins was the 35％ethanol. The methodological study on similarity and RSD in ARF HPLC fingerprint of three batches of samples cor-responded to related regulations. Conclusion The extraction and enrichment process of ARF is stable and repeatable.

To investigate the influence of G-CSF mobilization on functions of donor T lymphocyte subpopulation and acute graft-versus-host disease, peripheral blood samples of 20 healthy donors were collected before and after G-CSF mobilization. The whole blood was diluted with IMDM in ratio of 1:1 and then incubated with PMA + ionomycin + monensin at 37 degrees C, 5% CO2 for 4 hours. After being mobilized and stained, the IL-4, IFN-gamma and IL-2 positive cells were counted with three-color flow cytometry. The results showed that before G-CSF mobilization, the percentages of donor's CD3(+)IFN-gamma(+), CD4(+)IFN-gamma(+), CD8(+)IFN-gamma(+) T cells were 3.2% (0% - 45.9%), 1.3% (0% - 23.8%) and 1.5% (0% - 22.2%) respectively. The percentage of above mentioned cells in donor increased to 19.2% (0% - 53.9%), 9.5% (0% - 49.5%), 7.5% (0% - 38.1%) respectively after G-CSF mobilization. The IL-2 positive CD3(+), CD4(+) and CD8(+) T cell percentage in pre-G-CSF mobilized donors was 1.5% (0% - 31%), 0.8% (0% - 30.0%) and 0% (0% - 5.3%) respectively and subsequently increased to 25.7% (0% - 51%), 19.8% (0% - 39.7%), 4.6% (0% - 20.9%) respectively after G-CSF mobilization. The IL-4 positive T subpopulation did not increased significantly after G-CSF mobilization. In the early stage after peripheral blood stem cell transplantation, donor's Tc1 percentage in aGVHD group was significantly higher than that in non-aGVHD group. The morbidity of severe aGVHD in high Tc2 percentage group was significantly lower than that in low Tc2 percentage group. It is concluded that the donor's type I T cells increase after G-CSF mobilization, the Tc1 percentage of G-CSF mobilized donor is correlated with the occurrence of aGVHD in the early stage after HSCT, the percentage of Tc2 in donor is negatively correlated with aGVHD morbidity in recipients.
Adolescent ; Adult ; Female ; Graft vs Host Disease ; etiology ; Granulocyte Colony-Stimulating Factor ; adverse effects ; Hematopoietic Stem Cell Mobilization ; adverse effects ; methods ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Middle Aged ; Recombinant Proteins ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; immunology

Objective:To detect the colony number of bacteria, yeasts and molds in fermentation process of Pinelliae Rhizoma Fermentata (PRF), microbial flora species, and quantitatively analyze the dynamic changes of four dominant microorganisms at different fermentation time points of PRF, so as to provide experimental basis for exploring the processing mechanism of PRF. Method:According to Pharmaceutical Standard Preparation of Traditional Chinese Medicine Prescription of Ministry of Health of the People's Republic of China (the 10^th volume), PRF was processed. The samples at five different fermentation time points (0, 30, 60, 90, 120 h) of PRF were taken, the culturing, isolation and purification of bacteria, yeasts and molds were carried out with selective media, and the colonies were counted. Fluorescence quantitative polymerase chain reaction (PCR) technique was employed to conduct absolute quantification of Bacillus subtilis, Paecilomyces variotii, Byssochlamys spectabilis and Aspergillus niger. The recombinant plasmids of these 4 microorganisms were used as the standard substances, and the standard curves were prepared after dilution of multiple ratios, quantitative analysis was performed on these 4 microorganisms in five samples at different processing time points (0, 30, 60, 90, 120 h) of PRF. Result:During the fermentation process of PRF, the number of bacteria was low with smooth change, while molds and yeasts grew dramatically at the late stage of fermentation and reached 1×10⁶ CFU·mL^-1 at the end of fermentation. At 5 different fermentation time points, the copy numbers of Bacillus subtilis were 3.53×10⁵, 7.56×10⁴, 1.58×10⁵, 1.90×10⁶, 1.85×10⁶ copies·g^-1, the copy numbers of Paecilomyces variotii were 0, 0, 0, 3.45×10⁷, 4.15×10⁸ copies·g^-1, the copy numbers of Byssochlamys spectabilis were 0, 0, 0, 1.04×10⁸, 2.28×10⁸ copies·g^-1, the copy numbers of Aspergillus niger were 0, 0, 9.48×10⁵, 1.47×10⁶, 7.56×10⁶ copies·g^-1, respectively. Conclusion:The change trend of microflora in the fermentation process of PRF can be reflected by the dynamic change of four dominant microorganisms, and molds may play an important role in the processing of PRF. Fluorescence quantitative PCR technique has the advantages of rapidity, sensitivity, good repeatability and high specificity, it is suitable for exploring processing mechanism of PRF.

ObjectiveWearing protective masks for a long time causes a large number of frontline health care workers to suffer different degrees of pressure injury or facial skin rupture in response to COVID-19. This paper aims to analyze the occurrence characteristics and related factors of pressure injury related to devices, and provide the basis for taking countermeasures. Methods There was online investigation of skin injuries caused by wearing protective equipment in medical staff. Descriptive analysis was carried out on the occurrence characteristics of pressure-induced injury, and influencing factors were analyzed through logistic regression model. ResultsThere were a total of 2901 valid questionnaires. The results showed that the incidence of pressure injury caused by protective equipment was 26.34%, mainly in the bridge of the nose (20.41%), cheek (20.23%), auricle (17.82%) and forehead (8.86%). Multivariate Logistic regression model analysis showed that the major associated factors, which presented increasing risk, were sweating and dampness (OR=12.72, 95%CI 8.36-17.30), wearing level-3 protective equipment (OR=3.55, 95%CI 2.47-5.08), wearing level-2 protective equipment (OR=3.37, 95%CI 2.47-4.60), wearing time (OR=1.29, 95%CI 1.05~1.58) and occupation (OR=1.57, 95%CI 1.00~2.49). Conclusion There is the high incidence of pressure injuries to health care workers caused by protective equipment against COVID-19. The main risk factors for facial stress injury of medical staff are sweating and dampness, wearing level-3 and level-2 protective equipment and wearing time.

A high-content GABA was found in Sojae Semen Praeparatum(SSP), which is a famous traditional Chinese medicine and officially listed in Chinese Pharmacopoeia. To screen out and identify GABA-producing microbes from samples at different time points during the fermenting process of SSP, traditional microbiological methods combined with molecular biological methods were used to study the predominant GABA-producing microorganisms existing in the fermenting process of SSP. This study would lay a foundation for further studying the processing mechanism of SSP. The fermenting process of SSP was based on Chinese Pharmacopoeia(2010 edition), and samples were taken at different time points during the fermenting process of SSP. The bacteria and fungi from samples at different time points in the fermenting process of SSP were cultured, isolated and purified by selective medium, and dominant strains were selected. The dominant bacteria were cultured in the designated liquid medium to prepare the fermentation broths, and GABA in the fermentation broth was qualitatively screened out by thin-layer chromatography. The microbial fermentation broth with GABA spots in the primary screening was quantitatively detected by online pre-column derivatization and high performance liquid chromatography established in our laboratory. GABA-producing microorganisms were screened out from predominant strains, and their GABA contents in fermentation broth were determined. The DNA sequences of GABA-producing bacteria and fungi were amplified using 16S rDNA and 18S rDNA sequences by PCR respectively. The amplified products were sequenced, and the sequencing results were identified through NCBI homology comparison. Molecular biological identification was made by phylogenetic tree constructed by MEGA 7.0 software. Through the homology comparison of NCBI and the construction of phylogenetic tree by MEGA 7.0 software, nine GABA-producing microorganisms were screened out and identified in this study. They were Bacillus subtilis, Enterococcus faecium, E. avium, Aspergillus tamarii, A. flavus, A. niger, Cladosporium tenuissimum, Penicillium citrinum and Phanerochaete sordida respectively. For the first time, nine GABA-producing microorganisms were screened out and identified in the samples at different time points during the fermenting process of SSP in this study. The results indicated that multiple predominant GABA-producing microorganisms exist in the fermenting process of SSP and may play an important role in the formation of GABA.
Bacteria ; classification ; metabolism ; Chromatography, High Pressure Liquid ; Fermentation ; Fungi ; classification ; metabolism ; Phylogeny ; Seeds ; microbiology ; Soybeans ; microbiology ; gamma-Aminobutyric Acid ; biosynthesis

BACKGROUND: Pulmonary fibrosis is a respiratory disease caused by the proliferation of fibroblasts and accumulation of the extracellular matrix (ECM). It is known that the lung ECM is mainly composed of a three-dimensional fiber mesh filled with various high-molecular-weight proteins. However, the small-molecular-weight proteins in the lung ECM and their differences between normal and fibrotic lung ECM are largely unknown. METHODS: Healthy adult male Sprague-Dawley rats (Rattus norvegicus) weighing about 150 to 200 g were randomly divided into three groups using random number table: A, B, and C and each group contained five rats. The rats in Group A were administered a single intragastric (i.g.) dose of 500 μL of saline as control, and those in Groups B and C were administered a single i.g. dose of paraquat (PQ) dissolved in 500 μL of saline (20 mg/kg). After 2 weeks, the lungs of rats in Group B were harvested for histological observation, preparation of de-cellularized lung scaffolds, and proteomic analysis for small-molecular-weight proteins, and similar procedures were performed on Group C and A after 4 weeks. The differentially expressed small-molecular-weight proteins (DESMPs) between different groups and the subcellular locations were analyzed. RESULTS: Of the 1626 small-molecular-weight proteins identified, 1047 were quantifiable. There were 97 up-regulated and 45 down-regulated proteins in B vs. A, 274 up-regulated and 31 down-regulated proteins in C vs. A, and 237 up-regulated and 28 down-regulated proteins identified in C vs. B. Both the up-regulated and down-regulated proteins in the three comparisons were mainly distributed in single-organism processes and cellular processes within biological process, cell and organelle within cellular component, and binding within molecular function. Further, more up-regulated than down-regulated proteins were identified in most sub-cellular locations. The interactions of DESMPs identified in extracellular location in all comparisons showed that serum albumin (Alb) harbored the highest degree of node (25), followed by prolyl 4-hydroxylase beta polypeptide (12), integrin β1 (10), apolipoprotein A1 (9), and fibrinogen gamma chain (9). CONCLUSIONS Numerous PQ-induced DESMPs were identified in de-cellularized lungs of rats by high throughput proteomics analysis. The DESMPs between the control and treatment groups showed diversity in molecular functions, biological processes, and pathways. In addition, the interactions of extracellular DESMPs suggested that the extracellular proteins Alb, Itgb1, Apoa1, P4hb, and Fgg in ECM could be potentially used as biomarker candidates for pulmonary fibrosis. These results provided useful information and new insights regarding pulmonary fibrosis.