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Objective:To construct a plasmid for expression Mycobacterium tuberculosis ( Mtb) Rv3133c and to evaluate the immunogenicity of Rv3133c through population and mice experiments. Methods:The recombinant expression plasmid pPROEX-Rv3133c was constructed. The transformed E. coli BL21 (DE3) carrying expression plasmid was induced by IPTG to express the recombinant Rv3133c (rRv3133c). Western blot was used to identify the expressed protein. Whole-blood IFN-γ release assay (WBIA) was preformed to assess the immunogenicity of rRv3133c in Mtb-infected population. Antigen-specific antibodies in serum, Th1 type cytokines in splenocytes, functional T cell subset responses in splenocytes and the expression of cytokines at mRNA level in lung tissues were detected after immunizing mice subcutaneously with rRv3133c and adjuvant DC. Results:The rRv3133c was constructed and expressed successfully. Stimulation with rRv3133c promoted the production of IFN-γ in Mtb-infected population, especially in patients with latent tuberculosis infections. After immunizing mice with rRv3133c+ DC, the levels of IFN-γ, TNF-α and IL-2, the number of IFN-γ + TNF-α + CD4 + T cells in spleen and the expression of antigen-specific IFN-γ, TNF-α and iNOS at mRNA level in lung tissues were higher than those in BCG-immunized mice, but lower than those in BCG+ rRv3133c+ DC group. The serum IgG2a/IgG1 ratios in the rRv3133c+ DC group and the BCG+ rRv3133c+ DC group were greater than 1, and significantly higher than that of the BCG group. Conclusions:The rRv3133c had good immunogenicity and could induce strong Th1 immune response, suggesting that it was a potential candidate antigen for subunit vaccine against tuberculosis.
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OBJECTIVE To explore the effects of Achyranthis b identatae-Cynanchum otophyllum as core couplet medicinals of Zhengan xifeng decoction on oxidative stress of cerebral tissue in Parkinson ’s disease （PD）mice with syndrome of hyperactivity of liver-Yang. METHODS C57BL/6 mice were randomly divided into normal control group ，model group ，Zhengan xifeng decoction group，A. bidntatae group，C. otophyllum group and couplet medicinals of A. bidentatae-C. otophyllum group，with 10 mice in each group. PD model of hyperactivity of liver -Yang was established by intragastric administration of Aconitum carmichaelii decoction（4 g/kg） and intraperitoneal injection of 1-methyl-4-phenyl-1，2，3，6-tetrahydropyridine （25 mg/kg）. The behavioral changes and ultrastructure of substantia nigra neurons in mice were observed . The expressions of tyrosine hydroxylase （TH）positive neurons in substantia nigra were detected . The expressions of total antioxidant capacity （T-AOC），superoxide dismutase （SOD） and malondialdehyde（MDA）in substantia nigra as well as mRNA and protein expressions of nuclear factor -erythroid 2-related factor 2 （Nrf2），thioredoxin reductase 1（Trxr1），thioredoxin interacting protein （Txnip）were also determined . RESULTS Compared with model group ，PD behavior and ultrastructure of substantia nigra neurons were all improved in administration groups . The expressions of TH positive neurons ，T-AOC（except for C.otophyllum group）and SOD （except for C.otophyllum group），mRNA relative expression and protein expression levels of Nrf 2 and Trxr 1 were all increased significantly ；the expression of MDA （except for C.otophyllum group）and mRNA relative expression and protein expression levels of Txnip were all decreased significantly （P＜0.05）. Intervention effect of couplet medicinals of A. bidentatae-C. otophyllum group was better than that of A.bidntatae group and C. otophyllum group（P＜ 0.05），and the effect was similar to that of decoction group （P＞0.05）. CONCLUSIONS The couplet medicinals of A. bidentatae-C. otophyllum can inhibit the level of oxidative stress in the cerebral tissue of PD mice with hyperactivity of liver -Yang by targeting Nrf 2，and play a protective role on the brain neurons . Its effect is better than that of A.bidntatae and C.otophyllum，and it plays the same role as that of the formula in tonifying the kidney ，softening the liver and suppressing the Yang .
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Angucyclines/angucyclinones are a large group of polycyclic aromatic polyketides and their producers are widely distributed in nature. This family of natural products attracts great attention because of their diverse biological activities and unique chemical structures. With the development of synthetic biology and the exploitation of the actinomycetes from previously unexplored environments, angucyclines/angucyclinones-like natural products with new skeletons were continuously discovered, thus enriching the structural diversity of this family. In this review we summarize the new angucyclines/angucyclinones analogues discovered in the last decade (2010-2020) by using different strategies, such as changing cultivation conditions, genetic modification, genome mining, bioactivity-guided compound isolation, and fermentation of actinomycetes from underexplored environments. We also discuss the role of synthetic biology in the discovery and development of new compounds of the angucycline/angucyclinone family.
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Anthraquinones , Biological Products , Polyketides , StreptomycesABSTRACT
Objective To explore the advantages and characteristics of pathology experimental teaching based on WeChat public platform. Method Through the establishment ofhomogeneous pathol-ogy experiment platform—WeChat public platform, the students of clinical medicine major of Class 1 and 2 of Grade 2014 who participated in the pathological experiment course were divided into two groups. The experimental group (65 people) used the auxiliary teaching based on the WeChat public platform, and the control group (65 people) used the traditional teaching method. The students in the experimental group used the WeChat public platform in combination with pathology experimental teaching, pushing the experimental teaching by WeChat public platform, including the change of specimen and eye diseases under the micro-scope of typical pictures and videos, and at the same time pushing the typical disease image and the related text introduction, and auxiliary pathology experimental teaching. The effect of teaching was evaluated by student experimental examination and electronic questionnaire. The data were collated after the entry of SPSS 19.0, and the data comparison was performed by t test. Result The average score of the experimental group (14.80±0.24) was significantly higher than that of the traditional teaching group (13.79±0.33), and the difference was statistically significant (P=0.031). The experimental group students' evaluation on the learning of WeChat based public platform was higher than the control group's evaluation on traditional teaching in the aspects of learning interest, reducing learning pressure and feedback. Conclusion The application of WeChat public platform for the teaching of pathological experiments is feasible compared with the traditional teaching model and can improve the teaching effect effectively, and solve the problems of the pathology experiment teaching sample being insufficient or the teaching sample being not typical, which creates conditions for students to study independently and use specimens and pictures.
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Objective To observe the protective effect of the traditional Chinese medicine prescription, Jiu Nao Yi Zhi water extract, on human neuroblastoma SH-SY5Y cell line, its effect on expression of insulin signal transduction pathway, and to explore the related mechanisms.Methods SH-SY5Y cells cultured in vitro, were divided into control group, Jiu Nao Yi Zhi No.1 prescription group and No.3 prescription group.The doses were 0.0625 mg/mL, 0.125 mg/mL, 0.25 mg/mL, 0.5 mg/mL and 1 mg/mL.The thiazolyl blue ( MTT) metabolic rate of each group was determined.The dose of 0.125 mg/mL was chosen for cell immunofluorescence analysis, and to observe the expression of insulin receptor substrates-1 ( IRS-1 ) , cAMP response element binding protein ( CREB ) , and the factors of insulin signal transduction pathway.Results Compared with the control group, MTT metabolic rates of the Jiu Nao Yi Zhi groups were significantly increased (P<0.05), and the cell morphology was much better in those groups, cell body more plump, well-adherent and neurite extensions were observed.The expressions of IRS-1 and CREB were higher than that in the control group.Conclusions The traditional Chinese medicine prescription Jiu Nao Yi Zhi water extract can protect neurons by promoting nerve cell growth, and improving the expression of IRS-1 and CREB, the factors of insulin signal transduction pathway.
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The incidence of pediatric adenoid hypertrophy increases year by year. However, the treatment methods are limited. By now TCM treatment has showed significant clinical efficacy in treating pediatric adenoid hypertrophy. And phlegm could be one of the main pathogenic factors. By collecting relevant literature about TCM treatment for pediatric adenoid hypertrophy, this article summarized the thoughts of treating from phlegm based on the collection and analysis of classification of syndrome and treatment methods guided by the thoughts of treating from phlegm.
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Objective To explore the application effects of simulation diagnosis teaching in practical teaching of obstetrics and gynecology.Methods Students were divided into simulation teaching group(n=46) and traditional teaching group(n=46).Survey was conducted among simulation teaching group and traditional teaching group concerning clinical teaching of obstetrics and gynecology.Teaching effects were evaluated by questionnaire and examination.SPSS 13.0 statistics software was used to do the analysis.Enumeration data was analyzed by chi-square test while measurement data by t test.P＜0.05 stands for statistically significant difference.Results There were significant differences in improving students' clinical thinking and practical ability between two groups(P＜0.05).Satisfaction degree of simulation teaching group was significantly higher than that of traditional teaching group.Examination results of simulation teaching group was significantly higher than that of traditional teaching group(theoretical exam:P=0.001; operational exam:P=0.000).Conclusions Simulation diagnosis teaching can provide students more opportunities to practice and improve students' clinical thinking ability and clinical practice ability.
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The chemical constituents of Chinese red ginseng (Panax ginseng) were investigated. The chemical constituents were isolated and purified by silca gel, ODS, and Sephedex LH-20, column chromatography, and preparative HPLC. Their chemical structures were elucidated on the basis of physicochemical properties and spectra data. Fourteen compounds were isolated and identified as: notoginsenoside R2 (1), 20(S) -ginsenoside Rg3 (2), 20(R) -ginsenoside Rg3 (3), 20 (S)-ginsenoside Rg2 (4), 20(R) -ginsenosideRg2 (5), 20 (S)-ginsenoside Rh1 (6), 20(R) -ginsenoside Rh1 (7), ginsenoside Rh4 (8), -Ro (9), -Rb1 (10), -Rg1 (11), Re-(12), Rf (13), maltol (14). Compounds 1, 4, 6, were obtained from red ginseng for the first time. Compounds 2 and 3, 4 and 5-7 were enantiomers respectively, enantiomers 6 and 7 were isolated as monomer for the first time.
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Ginsenosides , Chemistry , Panax , Chemistry , StereoisomerismABSTRACT
Objective To develop urine AD7C-NTP diagnostic kit,analyze and evaluate its application value on AD.Methods Immunogenicity AD7C-NTP peptide fragments had synthesized by solidphase methods.The animals immunized to prepare antibodies.After matching screening.mouse antibody was uesed as coating antibody.biotin-labeled rabbit antibody wag used as testing antibody,and horseradish peroxidase was labeied with avidin.The urine AD7C-NTP ELISA detective method was established.The AD7C-NTP levels in morning urine samples of 121 AD patients and 118 age-matched controls were collected.Results AD7C-NTP antibodies were identified.Mouse anti-AD7C-NTP antibody titer in ELISAwas 1:8 000,and rabbit anti-AD7C-NTP antibody titer in ELISA was 1:32 000:WB was uesd to detect human brain specimens and there was a single band with molecular weight of 41 000.The lowst detection limit of ELISA methodology was 0.5μg/L The linear range was 0-10μg/L,normal reference value ≤1.5μg/J,the average recovery rate was 100.2%.The intra and inter of CV were 3.8%,4.5%,7.6%,6.8% respectively.The AD7C-NTP levels[2.25(0.43-8.62)μg/L]of urine in AD group was higher than those in contorl group[0.82(0.47-2.77)μg/L,P<0.01].The positive rates in AD group and control group were 89.3% and 15.3% respectively.The sensitivity Was 89.3%and specificity was 84.7%.Conclusions The animals are immunized with the self-designed synthetic peptide fragment to prepare AD7C-NTP antibodies successfully.The established ELISA method for detection of urine AD7C-NTP with high sensitivity,and precision can be used as an assistant examination in clinical diagnosis of AD.
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Objective To look for a small molecular neurotrophin which could promote production and/or differentiation of neural stem cells (NSCs). Methods 1.Embryo hippocampal NSCs were cultured in vitro . 2. The neurospheres were identified by antibodies of bromodeoxyuridine (BrdU), glial fibrillary acid protein (GFAP), microtubule associated protein 2 (MAP2) and Galactocerebroside (GalC). 3. The cells were divided into four groups, which were control group, FBS treated group, trans-sequence of APP 5-mer peptide treated group, APP 5-mer peptide treated group. The morphology of NSCs was observed in above four groups. 4. Cell counts, detection of clone information rate and diameter of clone were done to study the effect of APP 5-mer peptide on production of NSCs. 5. In addition, we also detected the MTT metablism rates in all groups. Results 1. The NSCs formed neurospheres and grew in floating. They were BrdU-positive. GFAP-positive, MAP2-positive and GalC-positive cells appeared after FBS were added into the medium. 2. The morphology of NSCs was not changed in APP 5-mer peptide treated group and trans-sequence group compared with the control group. 3. The cell number increased in APP 5-mer peptide treated group as compared with the control group. There were no apparent differences between the control group and the trans-sequence treated group. 4. The clone formation rate and diameters of neurospheres increased in APP 5-mer peptide treated group. 5. The MTT metabolism rate increased in APP 5-mer peptide treated group. Conclusion APP 5-mer peptide could promote the production of embryo hippocamal NSCs in vitro. APP 5-mer peptide doesn't promote the differentiation of embryo hippocamal NSCs in vitro.
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Objective To establish an ultraviolet-irradiation damage model in cultured fibroblasts derived from human skin and to explore the potential protective effects and mechanisms of amyloid precursor protein 17-met peptide (APP17-mer peptide) on the oxidative damage and collagen metabolism in cultured fibroblasts after ultraviolet irradiation. Methods Human dermal fibroblast cultures were established by outgrowth from foreskin biopsies of a healthy donor and were irradiated by a single exposure to ultraviolet rays and cultured in a series of concentrations of APP17-mer peptide (0, 20, 40, 80 μmol/L).The activity of fibroblasts was detected by the assay of MTT. The intracellular ROS level was measured with a confocal microscope. The expression of MMP-1 mRNA was analyzed real-time quantitatively following RT-PCR. Results Primary cultures of human skin fibroblasts were established from human foreskin in DMEM supplemented with 10 % fetal bovine serum. UV irradiation depressed cellular activity and increased intracellular level of ROS (P<0.05). 40μmol/L and 80μmol/L APP17-mer peptide increased the cellular activity in both UV irradiated fibroblasts and unirradiated fibroblasts (P<0.05), however,20 μmol/L did not show such protective effects (P>0. 05). 40μmol/L APP17-mer peptide could depress the level of ROS in irradiated libroblasts. A single exposure of fibroblasts to UV irradiation resulted in 1.78 foldup-regulation of MMP-1 mRNA compared with unirradiated sample, 40μmol/L and 80μmol/L APP17-mer peptide decreased the expression of MMP-1 mRNA (P<0.05 and P<0.01, respectively).Conclusion APP17-mer peptide can enhance cellular activity under UV-induced oxidative stress and in-hibit collagen degradation in fibroblasts irradiated with ultraviolet rays. Inhibition of ROS production may be involved in the protective mechanism of APP17 peptide.
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BACKGROUND: Amyloid β(Aβ) protein is the core of senile plaque.Being the toxic segment of Aβ, Aβ25-35 has been extensively applied in the experiments of recent years. The research in the past has verified that the self-prepared Chinese herb, fuzhisan can promote the survival of the cultured neural cells and probably acts on the treatment of Alzheimer disease (AD).OBJECTIVE: To study the resistance of fuzhisan to Aβ25-35 toxicity to cultured neural cells and the probable approaches.DESIGN: Repeated measurement based on the cells.SETTING: Department of neurology of a university hospital and key experimental room in brain aging in a university hospital.MATERIALS: The experiment was performed in Beijing Key Experimental Room in Brain Aging of Xuanwu Hospital from June 2002 to April 2003. Dopaminergic SH-SY5Y cell of neuroblastoma and Aβ25-35 were employed. Chinese herb, fuzhisan was decocted with mild fire and its upper clear solution was collected and prepared into storage solution at the concentration of 0. 5 g/mL. Antibody: Beijing Key Experimental Room in Brain Aging of Xuanwu Hospital prepared cAMP responsive element binding protein(CREB),Bcl-2 in B lymphatic leukaemia-2 genetic product and cytochrome C(CytC).METHODS: SH-SY5Y cell was incubated with Aβ25-35 of various doses alone or in combination with fuzhisan and was compared with blank control. MTF metabolic rate of cultured neural cells were determined under different incubation conditions. Western-blot method was used to measure the protein expression changes in incubation with fuzhisanalone, incubation with Aβ25-35 alone and the combination incubation, compared with the blank control.MAIN OUTCOME MEASURES: It was to study MTT metabolic rate in the comparison between each experimental group and the blank control and expressions of CREB, Bcl-2 and CytC relevant to survival/death of neural cells.RESULTS: Survival rate of SH-SY5Y cell was increased by 11.4% in incubation with fuzhisan alone. It was remarkably improved in incubation combining fu zhisan with Aβ25-35 as compared with Aβ25-35 alone. The expressions of CREB and Bcl-2 in Aβ25-35 group were decreased and were increased in fuzhisan group. CytC expression in cytoplasm was increased in Aβ25-35 group and was declined with fuzhisan incubation.CONCLUSION: Fuzhisan promotes the survival of cultured neural cells and its protection is still existed under Aβ25-35 injury. Fuzhisan brings such effects into play probably by the protein expressions relevant to survival/death of cells.
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BACKGROUND: It has been verified in the experiments over the past that the self-prepared Chinese herb, fuzhisan can retard natural aging in rats, suggesting that such drug acts on anti-aging.OBJECTIVE: To observe the effect of optimum effective concentration of nerve cell cultured with fuzhisan on morphological alternation of neuroblastoma SH-SY5Y cell.DESIGN: Repeated measurement.MATERIALS: The experiment was performed in Beijing Key Laboratory Room of Cerebral Aging of Xuanwu Hospital Affiliated to Capital University of Medical Sciences from June 2002 to April 2003. Self-prepared Chinese herb, fuzhisan [composed of 6 herbs, such as shichangbu (Rhizoma Acori Graminei), yuanzhi (Radix Polygalae), etc.] was co-developed by Prof.Wang De-shen from Department of Neurology of First Clinical Medical College of Harbin Medical University and Prof. Xu Xiao-yun from Department of Neurology of Shanghai Oriental Hospital. In addition, amyloid βprotein 25-35 segment and SH-SY5Y neuroblastoma were provided.METHODS: Fuzhisan of various concentrations were used for incubation of neuroblastoma SH-SY5Y cell. Thiazolyl blue (MTT), colorimetric method was used to determine the cell survival rate. Dose-effect relationship curve was drawn up to search optimum drug concentration. The cells cultured with 6-pore plate were divided into normal control, amyloid β-protein 25-35 25 μmol/L group, amyloid β-protein 25-35 25 μmol/L + fuzhisan 45×10-3 g/L groups and fuzhisan 45×10-3 g/L group. They were incubated for 24 hours to observe cell morphological alternation and determine neurosome area and axon length.ery group.length of every group: Those in amyloid β-protein 25-35 25 μmol/L group were decreased remarkably than the normal control [(505.5 ±122.36),(599.8 ±141.25) μm2; (26.0±13.97), (36.5 ±15.58) μm, (t =3.903,3.447, P=0.000)]. Those in fuzhisan 45×10-3g/L group and amyloid β-protein 25-35 25 μmol/L + fuzhisan 45×10-3 g/L group were increased remarkably than amyloid β-protein 25-35 25 μmol/L group [(918.3±178.34),(896.6 ±257.14), (505.5 ±122.36) μm2; (96.8 ±43.31), (88.3 ±30.23),(26.0±13.97) μm, (t=10.922, 14.172, P=-0.000)].CONCLUSION: With injury of amyloid β-protein 25-35, fuzhisan still enhances the survival of cultured nerve cell, manifested as promoting the increase of neurosome area and axonal extension.
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Objective To study the effect of pravastatin on the patients with chronic renal failure(CRF)and normal blood lipid. Methods 48 cases of CRF patients with normal blood lipid were randomly divided into two groups. 26 patients of group 1 were given pravastatin besides basic treatment. 22 patients of group 2 were only given basic treatment. The treatment was consecutive for one year. Blood lipid, serum creatinine, clearance rate of serum creatinine(Ccr), urinary retinol-binding protein(RBP) and protein amount in urine were tested respectively before treatment and one month, six months and one year after the treatment. Results Compared with the patients of group 2, blood lipid, urinary RBP and total urinary proteins and serum creatinne decreased, and Ccr increased in the patients of group 1. Conclusion Pravastatin can postpone the renal dysfunction of the CRF patients with nomal blood lipid.
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OBJECTIVE:To investigate the effects of different processed products of Paeonia lactifloria in Shaoyao gancao tang on the content of paeoniflorin in their use.METHODS:The content of paeoniflorin in Shaoyao gancao tang was determined by HPLC in which SHIMA-DCIS ODS C18 was used as chromatographic column with acetonitrile-0.1% phosphoric acid water solution(15∶85) used as a mobile phase at a flow rate of 1.0 mL?min-1.The detective wavelength was set at 230 nm.RESULTS:In Shaoyao gancao tang,the content of paeoniflorin in Radix Paeoniae Rubra was the highest,followed in descending order by Radix Paeoniae Alba Prepara cum Vini,Radix Paeoniae Alba Prepara Flava,Radix Paeoniae Alba sine Cortice and Radix Paeoniae Alba. CONCLUSION:The difference in the content of paeoniflorin appeared when Radix Paeoniae Rubra,Radix Paeoniae Alba Prepara cum Vini and Radix Paeoniae Alba respectively used in combination with Shaoyao gancao tang suggest that Radix Paeoniae Rubra,Radix Paeoniae Alba and its processed products should be used differently based on different syndrome in clinical use of Shaoyao gancao tang.
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Urinary microquantitative albumin (UALb), immunoglobulin G (UIgG) ?2-microglo-bulin (U?2m), and Tamm-Horsfall protein (THP) were measured in 36 chronic B hepatitis (CHB) and 23 liver cirrhosis (LC) patients with radioimmunoassay. 109 healthy normal subjects served as controls. The results showed that the excretion of UALb was significantly increased in 17 patients. Out of 17 patients,the excretion of UIgG in 5 and U?2m in 16 were increased and the excretion of THP in 9 patients were decreased. Compared with normal controls there was a significant difference (P