[Extraction of Extracelluar Vesicles Derived from Mycobacterium tuberculosis and Their Effect on the Production of Reactive Oxygen Species and Expression of Inflammatory Factors in Mouse Bone Marrow-Derived Dendritic Cells].

Objective: To isolate extracellular vesicles (EVs) from Mycobacterium tuberculosis ( Mtb), to examine their morphology, particle size, and distribution, to study the effect of EVs derived from Mtb ( Mtb-EVs) on intracellular reactive oxygen species (ROS) production and cytokine secretion in dendritic cells (DCs), and to make preliminary exploration of Mtb-EVs' effect on the immune regulation of DCs. Methods: Mtb-EVs were obtained by ultrafiltration concentration and the protein concentration was determined by BCA assay. The morphology of Mtb-EVs was observed through negative staining electron microscopy (EM). The particle size distribution and concentration of Mtb-EVs were determined by nanoparticle tracking analysis (NTA). Mouse bone marrow was isolated through sterile procedures and mice myeloid DCs were induced and amplified by the combined use of recombinant mouse granulocyte-macrophage colony-stimulating factor (rm GM-CSF) and recombinant mouse interleukin-4 (rm IL-4). Then, morphological and immunophenotypic characterization was performed. After that, the DCs were treated with Mtb-EVs at different concentrations and CCK-8 assay was done to measure their effect on the survival rate of DCs and to identify the appropriate stimulation concentration for subsequent experimental procedures. The intracellular ROS levels of DCs were evaluated with DCFH-DA fluorescence probe and the cytokine secretion of DCs was determined by ELISA. Results: EM observation showed that Mtb-EVs isolated by ultrafiltration concentration were spherical vesicles of varied sizes, all being approximately 100 nm in diameter and displaying typical morphology. NTA results from NanoSight nanoparticle tracker showed that the peak particle size was 98.5 nm, that the average particle size was 110.2 nm, and that the particle size was mainly distributed between 68.4-155.7 nm. Mtb-EVs that were smaller than 250 nm accounted for 98.39% of the total. Mouse myeloid DCs directionally induced and amplified in vitro displayed typical DC phenotype and morphology, and the purity exceeded 85%. EM verified the abundance of microvilli and radial protuberance on the surface of DCs, which had uniform cytoplasm and clear nuclear membrane. Loaded with Mtb-EVs at different concentrations, including 10 2, 10 3, and 10 4 particles/cell, the DCs had significantly upregulated levels of intracellular ROS ( P<0.05). In addition, Mtb-EVs induced the release of IL-1ß and IL-6 in a dose-dependent manner ( P<0.05). Conclusion: We established in the study a technical process for the extraction of Mtb-EVs by ultrafiltration concentration and obtained Mtb-EVs with sound morphology, high purity, and concentrated particle size distribution. Furthermore, Mtb-EVs can upregulate the intracellular ROS level in DCs and induce the release of IL-1ß and IL-6 in a dose-dependent manner.

[Analysis of the Characteristics of the Outcomes of Job-Transfer Subspecialty Training Program in Pediatrics in Sichuan].

Objective: To discuss the characteristics of physician trainee outcomes after completion of the job-transfer subspecialty training in pediatrics, a program designed to increase the number of pediatricians, in Sichuan Province and to provide countermeasures for alleviating the shortage of pediatricians. Methods: We collected with questionnaire surveys information on changes in the workload and salaries experienced by physicians who completed the job-transfer subspecialty training program in pediatrics between February 2017 and May 2020 in Sichuan Province. Then, we compared the characteristics of physicians who successful became pediatricians and those who did no. Results: A total of 208 physicians completed the job-transfer subspecialty training program in pediatrics. Among them, 178, accounting for 85.6%, completed the questionnaire survey, and 120, accounting for 67.4%, had a background in other subspecialties than pediatrics. The majority (>90%) of physicians who participated in the training program came from secondary or lower levels of hospitals from the cities and prefectures all over Sichuan Province. In this study, we found that the rate of successful job transfer from being a physician to being a pediatrician in Sichuan Province in the past four years was 85.0% (102/120), with the year-by-year results being 88.2% (15/17) in 2017, 72.7% (16/22) in 2018, 86.7% (39/45) in 2019, and 94.% (32/34) in 2020. There was no significant difference between physicians who had successful job transfer and became pediatricians and those who failed to do so in terms of gender, age, hospital level, specialization prior to the job transfer, whether or not the hospital had a pediatrics department, amount of support for the pediatrics department, whether or not the physician was working at a new hospital after the job transfer, salaries, and changes of responsibilities during COVID-19 (all P>0.05). There was significant difference in the change of workload after completion of the training program between physicians who had successful job transfer and became pediatricians and those who failed to do so ( χ 2=9.037, P=0.003), and 78.4% of the trainees stated that their workload had increased after the job transfer. There was a moderate correlation between successful job transfer and changes in workload after the transfer (|Phi[ψ] |=0.729). Conclusions: The policy of government-supported job-transfer subspecialty training in pediatrics has played an active and important role in the swift resolution of the shortage of pediatricians. However, finding the root cause of and addressing the problem of the overwhelming workload of pediatricians remain challenging issues to be resolved.

[Immune Modulatory Effect of Outer Membrane Vesicles Derived from Salmonella on Mouse Bone Marrow-Derived Dendritic Cells].

OBJECTIVE: To study the effect of outer membrane vesicles (OMVs) derived from Salmonella typhimurium (ST) on the ultrastructural features and immune function of dendritic cells (DC). METHODS: Mice bone marrow cells were collected aseptically, and myeloid DC were generated by the combined induction and amplification with recombinant mouse granulocyte-macrophage colony-stimulating factor (GM-CSF) and recombinant mouse interleukin-4 (rm IL-4). Cell morphology was observed under inverted phase contrast microscope and the phenotype was identified with flow cytometry. ST-OMVs were isolated through ultracentrifugation. The survival rate of DC was assessed with CCK-8 assay, and the stimulus concentration of OMVs was henceforth determined. The ultrastructural characteristics of DC loaded with OMVs were observed with transmission electron microscopy. The cytokine secretion, surface molecule expression and phagocytic capacity of DC were examined with flow cytometry. RESULTS: The DC induced and amplified in vitro displayed typical DC phenotype in morphological analysis and the purity of DC exceeded 85%. Transmission electron microscopy showed that there were large numbers of protrusions on the cell surface. After stimulation with ST-OMVs, it was observed that the dendritic structures on the surface of DC were reduced and a large number of phagolysosomes were found in the cytoplasm. In addition, increased numbers of mitochondria, swelling and typical apoptosis were observed. After treatment with ST-OMVs at 5 µg/mL and 10 µg/mL, the secretion of tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) of DC increased significantly ( P<0.05). Furthermore, the immature DC could differentiate into mature DCs after stimulation with ST-OMVs, which were characterized by a decrease in phagocytic capacity ( P<0.05) and an upregulation of phenotypic markers ( P<0.05). CONCLUSION: ST-OMVs can stimulate DC to produce TNF-α and IL-1ß and promote DC maturation and antigen presentation.

[Proteomics analysis of erythrocyte membrane in rats with high altitude polycythemia before and after intervention with salidroside].

To investigate the effect of salidroside on the proteomics of erythrocyte membrane in high altitude erythrocytosis(HAPC) rats, in order to explore the mechanism of salidroside in improving HAPC based on the proteomics analysis. First, HPAC rat models were established, and 16 rats were randomly divided into HAPC model group and salidroside(100 mg·kg~(-1)) treatment group(8 rats per group). Saline was administered to the HAPC model group, while salidroside treatment group was given 100 mg·kg~(-1) salidroside once a day. After continuous oral administration with salidroside for 40 days(once a day), blood was collected from the femoral artery to obtain total red blood cell membrane proteins. Two-dimensional electrophoresis was used to separate total proteins. The two-dimensional electrophoresis of erythrocyte membrane proteins was analyzed before and after salidroside intervention, and the proteins with significant differences were identified by mass spectrometry. Finally, biological functions were analyzed using bioinformatics. A two-dimensional electrophoresis method was used to establish a protein expression profile with a high resolution and reproducibility of erythrocyte membranes in HAPC rats. Salidroside treatment significantly changed 18 protein spots in the 2-DE map of erythrocyte membranes, of which 13 proteins were up-regulated and 5 proteins were down-regulated. Eight differential proteins were successfully identified by mass spectrometry. Moreover, bioinformatics analysis found that these differential proteins were involved in such biological processes as oxidative stress, redox, and peroxisome pathway, which are mainly associated with peroxisome and MAPK signaling pathways. Therefore, salidroside could significantly change the expressions of erythrocyte membrane proteins in HAPC rats. Eight differential proteins were identified by a proteomic-based approach. The differential proteins were involved in such biological processes as oxidative stress, redox, peroxisome pathway.

Development and validation of the NCGG-FAT Chinese version for community-dwelling older Taiwanese.

AIM: To evaluate the reliability and validity of the National Center for Geriatrics and Gerontology functional assessment tool (NCGG-FAT) Chinese version among community-dwelling older Taiwanese. METHOD: In total, 40 community-living older adults aged ≥65 years with intact global cognitive function (Mini-Mental State Examination ≥24) were enrolled and received a neuropsychological assessment twice using the computerized NCGG-FAT Chinese version, with an interval of 30 days to examine test-retest reliability. Conventional neurocognitive assessments were performed for all study participants within a week after the first administration of the NCGG-FAT Chinese version to determine validity. Intraclass correlation coefficients (ICC) were employed to assess test-retest reliability, and the Pearson correlation coefficient evaluated the validity. RESULTS: In total, 40 participants aged 69.8 ± 3.9 years with a mean education of 11.1 ± 4.2 years and MMSE of 28.5 ± 1.8 were enrolled. The Pearson correlation coefficient showed moderate-to-high validity between the conventional neurocognitive assessments and the NCGG-FAT Chinese version components (r = 0.509-0.606, P < 0.01 for all components). High reliability was also identified in the word recognition (immediate) score (ICC = 0.833, P < 0.001), Trail Making Tests part B (ICC = 0.709, P < 0.001) and Symbol Digit Substitution Task score (ICC = 0.850, P < 0.001), whereas word recall (delayed) score and Trail Making Tests part A showed moderate test-retest reliability. CONCLUSIONS: The NCGG-FAT Chinese version is a valid and reliable instrument to assess multiple dimensions of neurocognitive function of community-living Taiwanese, which may facilitate better community-based screening and intervention programs, particularly for international comparisons. Geriatr Gerontol Int 2020; 20: 1171-1176.

COLEC12 regulates apoptosis of osteosarcoma through Toll-like receptor 4-activated inflammation.

OBJECTIVE: To investigate the role of COLEC12 in osteosarcoma and observe the relationship between COLEC12 knockdown and the inflammation of osteosarcoma. Then, further explore whether the process is regulated by TLR4. METHOD: GEPIA and TCGA systems were used to predict the potential function of COLEC12. Western blot and RT-PCR were used to analyze the protein expression, or mRNA level, of COLEC12 in different tissue or cell lines. The occurrence and development of osteosarcoma were observed by using COLEC12 knockdown lentivirus. The inflammation indexes of osteosarcoma, in vitro and in vivo, were explored. TLR4 knockdown lentivirus was applied to the relationship between COLEC12 and TLR4. RESULTS: COLEC12 expression in SARC tumor tissue was higher than in normal, and a high expression of COLEC12 in SARC patients had a worse prognostic outcome. Pairwise gene correlation analysis revealed a potential relationship between COLEC12 and TLR4. The COLEC12 expression and mRNA level in the tumor or Saos-2 cells were increased. COLEC12 knockdown lentivirus could inhibit osteosarcoma development, in vivo and vitro, through reducing tumor volume and weight, weakening tumor proliferation, migration, and invasion, and enhancing apoptosis. Furthermore, COLEC12 knockdown could increase inflammation of osteosarcoma, in vivo and in vitro, through inducing myeloperoxidase (MPO), TLR4, NF-κB, and C3, and expression of related inflammatory factors. Finally, TLR4 knockdown lentivirus inhibits the progress of inflammation after COLEC12 regulation, in vivo and vitro. CONCLUSION: COLEC12 may be able to regulate apoptosis and inflammation of osteosarcoma, and TLR4 may be the downstream target factor of COLEC12 in inflammation.

Antiviral effects of selected IMPDH and DHODH inhibitors against foot and mouth disease virus.

Foot-and-mouth disease virus (FMDV) is an important pathogen that affects livestock breeding and causes huge economic losses worldwide. Currently, the development of antiviral agents to combat FMDV infection at the early stages is being explored. As viral replication critically depends on the host for nucleoside supply, host enzymes involved in nucleotides biosynthesis may represent potential targets for the development of antiviral agents. In the present study, the effects of IMP dehydrogenase (AVN-944 and mycophenolate mofetil) and dihydroorotate dehydrogenase (teriflunomide) inhibitors were evaluated both in vitro and in vivo. The results revealed that these compounds were effective in suppressing FMDV (O/MY98/BY/2010 and A/GD/MM/2013) infection. With regard to the antiviral mechanism, time-of-addition experiments revealed that these compounds were effective when added at the early stages of viral lifecycle (0-8 h post infection). However, exogenous guanosine/uridine eliminated the antiviral activity of these compounds. Importantly, treatment AVN-944 and teriflunomide significantly improved the survival of mice that were subcutaneously treated with FMDV. Together, the results of the present study indicate the broad-spectrum activities of anti-FMDV agents targeting IMP dehydrogenase or dihydroorotate dehydrogenase, which could be useful in developing strategies to prevent FMD.

Thermoacoustic Tomography of In Vivo Human Finger Joints.

OBJECTIVE: the purpose of this study was to demonstrate the potential of thermoacoustic tomography (TAT) to reveal anatomic structures of in vivo human finger joints. METHODS: all the participating volunteers provided written informed consent. Eight healthy middle and index fingers from five volunteers were imaged in vivo by our TAT imaging system. Axial T1-weighted MR imaging (3.0 T) was used to validate the TAT findings. Comparative analyses between TAT and MRI images were performed in two dimension for all the fingers imaged. Three-dimensional (3-D) images and animations were also obtained for some of the fingers thermoacoustically scanned. RESULTS: various intra- and extra-articular tissues were identified in TAT images in high fidelity. These TAT images matched well with the MRI images. Both the 3-D images and animations effectively displayed the extension and transformation of the entire finger along the axis. CONCLUSION: TAT can noninvasively visualize anatomic structures of the finger joints based on the electrical properties of the joint tissues. The results obtained indicate that TAT may have the potential to contribute to the detection of joint and bone diseases. SIGNIFICANCE: this study represents the first for TAT of in vivo human joints and fingers. This study reveals that TAT can effectively recover both soft and hard tissues of the healthy interphalangeal joints, which provides a foundation for its clinical application to detection and diagnosis of joint and bone diseases.

[Risk analysis of imported schistosomiasis in Shanghai City from 2005 to 2015].

OBJECTIVE: To understand the risk situation of imported schistosomiasis and its main influencing factors in Shanghai City from 2005 to 2015. METHODS: A retrospective survey was performed, and the database was established based on the data collection of imported schistosomiasis, Oncomelania hupensis snail situation and mobile population in Shanghai City from 2005 to 2015. RESULTS: From 2005 to 2015, the mobile population in Shanghai City increased by 123.92%, which reached about 9 816 500, and 54.70% of them were located in the inner suburban districts. The accumulated areas with snails of 7.13 hm2 were found in 16 towns of 4 outer suburbs (excluding Chongming District). A total of 23 cases of imported schistosomiasis were found in Shanghai City, and the number of the imported schistosomiasis cases was on the downward trend (rs = -0.782, P = 0.004). From 2005 to 2009, 11 out of 13 imported schistosomiasis cases (84.62%) were distributed in the inner suburban districts where no O. hupensis snails were found at the same time. From 2010 to 2015, 9 out of 10 imported schistosomiasis cases (90%) were distributed in the outer suburban districts where O. hupensis snails were found at the same time. CONCLUSIONS: The number of imported schistosomiasis cases in Shanghai is on the downward trend. However, the threat of imported schistosomiasis to the prevention and control of schistosomiasis in Shanghai City should not be ignored.

[Comparative analysis of malaria detection ability of laboratories in Shanghai City from 2012 to 2015].

OBJECTIVE: To compare the application effects of three methods, namely microscopic examination, antigen detection (RDT) and nucleic acid test (PCR) in malaria detection between municipal and districts/counties centers for disease control and prevention in Shanghai, and analyze the malaria detection ability of the laboratories in Shanghai. METHODS: The blood smears, whole blood samples, case review confirmation records and case data of malaria cases and suspected cases in Shanghai from 2012 to 2015 were collected by Shanghai Municipal Center for Disease Control and Prevention, and the detection results were analyzed and compared. RESULTS: A total of 212 samples with complete data were submitted by all districts (counties) in Shanghai from 2012 to 2015, the samples submitted by Jinshan Districts were the most (41.98%), and among the first diagnosis hospitals, those submitted by the tertiary hospitals were the most (82.07%). The submitted samples in the whole year were increased gradually from January to October. All the 212 samples were detected by three methods (the microscopic examination, RDT and PCR) in the laboratory of Shanghai Municipal Center for Disease Control and Prevention, and 167 were tested and confirmed comprehensively as positives, accounting for 78.77%, and 45 were confirmed as negatives, accounting for 21.23%. The samples were detected by the method of microscopy and domestic RDT in the laboratories of the centers for disease control and prevention at district/county level, totally 153 were tested as positives, accounting for 72.17%, 41 were unclassified, accounting for 19.34%, 53 were negative, accounting for 25.00%, and 6 were undetected, accounting for 2.83%. The coincidence of microscopic examination between the report hospitals and the centers for disease control and prevention at district/county level was 78.16%, and the coincidence between centers for disease control and prevention at district/county level and municipal level was 93.20%. The utilization rate of RDT in the laboratory of district/county level was 73.58%. The coincidence of RDT tests between those domestic and imported was 93.59%. Compared with the detection results by municipal center for disease control and prevention, 37 samples were misjudged by the laboratories of district/county level. Almost all (99.37%) of the confirmed malaria cases were imported overseas, including Africa (85.44%), Asia (13.92%) and America (0.63%). CONCLUSIONS: The surveillance after malaria elimination in Shanghai should be carried out by combining with different detection methods and resource integration.

Clinical, Pathological, and Prognostic Characteristics of Glomerulonephritis Related to Staphylococcal Infection.

Staphylococcal infection has become a common cause of postinfectious glomerulonephritis in the past 3 decades. Because few investigations focus on this disease, the demographics and clinicopathological features of glomerulonephritis related to staphylococcal infection are not well characterized. We conducted a pooled analysis of published literature in electronic databases and analyzed the clinical features, laboratory findings, and histopathological changes. The patients were divided into 4 groups based on their prognosis: remission, persistent renal dysfunction, end-stage renal disease (ESRD), or death. A logistic regression model was used to identify the determinants of disease outcome. A total of 83 (64 men) patients with glomerulonephritis related to staphylococcal infection from 31 reports were analyzed. The mean age was 58 years (58 ±â€Š17). Majority of the reports originated from Taiwan, Japan, and the United States. Clinical characteristics of the cases were hematuria (82/83), proteinuria (78/83), and acute kidney injury (75/83). Visceral abscesses (26/83) and skin infections (24/83) were the common sites of infection. Methicillin-resistant Staphylococcus aureus was the most common pathogen. The dominant or codominant deposition of IgA or C3 along the glomeruli was an important feature identified by immunofluorescence. There were 19 patients (22.9%) that progressed to dialysis-dependent ESRD. Twelve patients (14.5%) died. A univariate regression analysis indicated that diabetes mellitus (DM) (odds ratio [OR] 2.96; 95% confidence interval [CI] 1.03-8.48; P = 0.04) and age (OR 4.80; 95% CI 1.84-12.53; P = 0.001) were risk factors for ESRD or death. A multivariate regression analysis also revealed that age (OR 4.90; 95% CI 1.82-13.18; P = 0.002) and DM (OR 3.07; 95% CI 0.98-9.59; P = 0.05) were independent risk factors for unfavorable prognosis. Glomerulonephritis related to staphylococcal infection has different features than typical postinfectious glomerulonephritis. The diagnosis of glomerulonephritis related to staphylococcal infection relies on immunofluorescence and electron microscopy findings. Age and DM are independent risk factors of poor prognosis for glomerulonephritis related to staphylococcal infection.

[Correlation between malaria epidemic incidence and some natural and social factors in high incidence areas of Shandong Province].

OBJECTIVE: To explore the association between malaria epidemic situation and some natural and social factors in high-incidence areas of Shandong Province, so as to provide evidences for malaria elimination in these areas. METHODS: Twenty towns of 10 counties (cities, districts,) in the high incidence areas of malaria in Shandong Province were selected as the study sites, and the residents in the study sites were investigated by questionnaires with one household as a unit, so as to understand the related natural and social factors. In addition, the malaria epidemic data in the study sites from 2006 to 2010 were collected, and the correlation between these factors and the epidemic situation of malaria was analyzed by Spearman rank correlation and multiple stepwise regression. RESULTS: The square root of malaria incidence rate (Y) was negatively related to the rate of households using insecticide (X3), and the rate of households using screen doors and windows (X4) (both P < 0.05), but was positively related to the rate of housing surrounding water environment and exposure ratio (X6) (both P < 0.05). The regression equation established was Y = 0.032X5 + 0.048X6-0.495, R2 = 0.973. CONCLUSIONS: Malaria incidence is obviously associated with some natural and social factors. The measures such as clearing the breeding place of mosquito, protecting the exposure population at nightfall, as well as using door-window screen and repellents correctly, can effectively control malaria.

[CT axial imaging of the iliolumbar ligament and its significance on locating lumbosacral vertebral segments].

OBJECTIVE: To study the CT axial manifestations of iliolumbar ligament(ILL) and discusses its clinical effects on locating lumbosacral vertebral segments. METHODS: From May 2008 to March 2010, 706 adult patients diagnosed lumbar disc disease were performed with axial scans by single slice helical CT. Among the patients, 436 patients were male and 270 patients were female, ranging in age from 25 to 82 years, the median age was 44 years, 78 cases with lumbosacral transitional vertebrae (LSTV) were verified by X-radiography or fluoroscopy. The morphology, origin and insertion, courses of ILL and the relationship of ligament and spinal segments on axial plane images were used to study. The location method of spinal segments by ILL was compared with the other four location methods on CT. RESULTS: Of the 628 cases with normal lumbosacral segmentations sides of ligament, the main part of ILL originated from L5 transverse processes and terminated at the iliac crest, the morphological characters were divided into two types: double band (71.8%, 451/628) and single band (28.2%, 177/628). The tiny branches from posterior and outside edge of L4, lumbar disc were seen simultaneity in 3 cases. The ILL of 78 cases with LSTV all also originated from L5 transverse processes. Using ILL as a marker of the L5 vertebral level, 78 cases with LSTV were correctly numbered, the accuracy rate was higher than the other location methods, there was statistical significance between the location method by ILL and the location method by iliac crest (P < 0.05). CONCLUSION: The main part of ILL originates from L5 transverse processes, the anatomic location is relatively steady and can be clearly displayed on axial CT, which can be used as a measure in the idenlification of LSTV in clinical practice, it is worthy to be applied widely in basic-level hospitals.

Genetic transformation of Gentiana macrophylla with Agrobacterium rhizogenes: growth and production of secoiridoid glucoside gentiopicroside in transformed hairy root cultures.

Hairy root cultures of Gentiana macrophylla were established by infecting the different explants four Agrobacterium rhizogenes strains namely A(4)GUS, R1000, LBA 9402 and ATCC11325, and hairy root lines were established with A. rhizogenes strain R1000 in 1/2 MS + B(5) medium. Initially, 42 independent hairy root clones were maintained and seven clones belongs to different category were evaluated for growth, morphology, integration and expression of Ri T-DNA genes, and alkaloid contents in dry root samples. On the basis of total root elongation, lateral root density and biomass accumulation on solid media, hairy root clones were separated into three categories. PCR and Southern hybridization analysis revealed both left and right T-DNA integration in the root clones and RT-PCR analysis confirmed the expression of hairy root inducible gene. GUS assay was also performed to confirm the integration of left T-DNA. The accumulation of considerable amounts of the root-specific secoiridoid glucosides gentiopicroside was observed in GM1 (T +/L and T +/R) and the GM2 (T +/L and T -/R DNA) type clones in considerably higher amount whether as two T -/L but T +/R callus-type clones (GM3) accumulated much less or only very negligible amounts of gentiopicroside. Out of four media composition the 1/2 MS + B(5) vitamin media was found most suitable. We found that initial establishment of root cultures largely depends on root:media ratio. Maximum growth rate was recorded in 1:50 root:media ratio. The maximum biomass in terms of fresh weight (33-fold) was achieved in 1/2 MS + B(5) media composition after 35 days in comparison to sixfold increase in control. The biomass increase was most abundant maximum from 15 to 30 days. Influence of A. rhizogenes strains and Ri plasmid of hairy root induction, the possible role of the T(L)-DNA and T(R)-DNA genes on growth pattern of hairy root, initial root inoculum:media ratio and effect of media composition is discussed.

Homing efficiency and hematopoietic reconstitution of bone marrow-derived stroma cells expanded by recombinant human macrophage-colony stimulating factor in vitro.

The increasing recognition of the properties of marrow stromal cells has spawned a major switch in our perception of their nature and the potential therapeutic applications that have been envisioned and implemented. Yet, several aspects of bone marrow stromal cell biology remain in question. This report describes the ability of recombinant human macrophage colony-stimulating factor (rhM-CSF) to maintain proliferation and differentiation of bone marrow stromal cells ex vivo. Our results demonstrated that M-CSF was essential for proliferation and differentiation of bone marrow-derived stromal cells and exerted its effects in a dose-dependent manner. The number of colony-forming unit (CFU) fibroblasts increased by 25% after incubation with rhM-CSF. In vitro expanded bone marrow stromal cells were easy to passage and differentiated to adipocyte and chondroblast cells under appropriate culture conditions. Furthermore, these expanded stromal cells to support CD34+ hematopoietic stem cells, as demonstrated by their ability to form CFU-Mix, burst-forming units-erythroid, and CFU-granulocyte macrophage colonies after 3 weeks of culture. The homing efficiency of in vitro expanded or fresh isolated bone marrow-derived stromal cells, which were labeled with carboxy fluorescein diacetate succinimidyl ester, to bone marrow was also investigated. Homing assays demonstrated that freshly isolated CD45+-depleted bone marrow cells were able to home to bone marrow in a dose-dependent manner, although some cells were found in the spleen, liver, and lung. However, their ability to home was dramatically reduced with culture time and was completely lost after five to seven passages in vitro. Animal studies showed that freshly isolated or rhM-CSF-induced bone marrow stromal cells promoted hematopoietic reconstitution in lethally irradiated mice. The ability to easily expand human stromal cells, which support survival and proliferation of CD34+ cells, has many important clinical applications for hematopoietic disorders.

The studies on radiological limits of color-glazed tiles used in home decoration.

This study was carried out to lay down the radiological limits of color-glazed tiles used in home decoration. The activity concentrations of various end products and raw materials as well as processed materials were measured using gamma spectroscopy. 222Rn exhalation rates from the surface of color-glazed tiles were measured using charcoal canister method. Levels of exposure to alpha ray and beta ray from the surface of glazed tiles were measured by surface alpha and beta contaminant instrument. The results show a great difference between the radioactive levels of the surface glaze and the matrix of color-glazed tiles. The 222Rn exhalation rates from the surface of color-glazed tiles are in the range of 10(-3) - 10(-4) Bq m(-2) s(-1). The concentrations of some natural radionuclides in glaze exceed the exempt limits of International Basic Safety Standards for Protection against Ionizing Radiation Sources (IAEA 1997). The limits for color-glazed tiles were deduced according to these data and the theory of UNSCEAR (1993). The activity concentration of 226Ra (Bq kg(-1)) in the glaze of color-glazed tiles should be in the range of A(226Ra) < or = 1,000 (Bq kg(-1)), and, at the same time, the specific activity of the natural radionuclides (Bq kg(-1)) should be in the range of A(232Th)/230 + A(226Ra)/310 + A(40K)/3,500 < or = 1.