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Objective: To establish and optimize PCR methods for the gene encoding of Clostridium perfringens β2 toxin (cpb2) and atypical-cpb2 (aty-cpb2), analyze the epidemiological characteristics and genetic polymorphism of the cpb2 of Clostridium perfringens in 9 Chinese areas from 2016 to 2021. Methods: The cpb2 of 188 Clostridium perfringens strains were examined by PCR; the cpb2 sequences were acquired by whole-genome sequencing to analyze the genetic polymorphism. Using Mega 11 and the Makeblastdb tool, a phylogenetic tree, and cpb2-library based on 110 strains carrying the cpb2 were produced. Using the Blastn technique, a comparison was made to discover sequence similarity between consensus-cpb2 (con-cpb2) and aty-cpb2. Results: The specificity of PCR assay for the cpb2 and aty-cpb2 was verified. The PCR results for cpb2 amplification were highly consistent with the whole-genome sequencing approach (Kappa=0.946, P<0.001). A total of 107 strains from nine regions in China carried cpb2, 94 types A strains carried aty-cpb2, 6 types A strains carried con-cpb2, and 7 types F strains carried aty-cpb2. The nucleotide sequence similarity between the two coding genes was 68.97%-70.97%, and the similarity between the same coding genes was 98.00%-100.00%. Conclusions: In this study, a specific PCR method for cpb2 toxin was developed, and the previous PCR method for detecting aty-cpb2 was improved. aty-cpb2 is the primary gene encoding of β2 toxin. There is a significant nucleotide sequence variance between the various cpb2 genotypes.
Subject(s)
Humans , Clostridium perfringens/genetics , Clostridium Infections , Bacterial Toxins/genetics , Phylogeny , Polymerase Chain Reaction , Polymorphism, GeneticABSTRACT
@#To improve the transduction efficiency of recombinant adeno-associated virus (rAAV) in NK92 cells, the number of cells, concentration of IL-2 in the medium, and serotype and dosage of rAAV were explored to optimize cell state and viral transfection conditions.Then, zinc chloride (ZnCl2), chloroquine, polyvinyl alcohol (PVA) and genistein with different concentration were added separately during transfection to further improve the viral transduction efficiency.The results showed that, at cell number of 5 × 105, the expression efficiency of enhanced green fluorescent protein (EGFP) was relatively high.When the IL-2 concentration was 1 000 IU/mL, NK92 cells were most suitable for virus transfection. The transduction efficiency of different serotypes of rAAV in NK92 cells was rAAV6, rAAV2 and rAAV9 in descending order.Pretreatment of NK92 cells with genistein could significantly increase the viral transduction efficiency, while the addition of other reagents had no significant effect.Through the optimization of the above conditions, the transduction efficiency of rAAV to NK92 cells could be significantly improved, which provided evidence for functional genetic modification of NK92 cells by rAAV.
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With the increasing aging population, the incidence of wet age-related macular degeneration(wARMD)is gradually rising. The formation of neovascularization leads to recurrent hemorrhage in the macular region, which is one of the main causes of blindness in the elderly. Currently, the primary clinical treatment for wARMD is intravitreal injection of anti-vascular endothelial growth factor(VEGF)drugs. However, there are still some patients who have poor or no response to anti-VEGF drugs, resulting in suboptimal or ineffective clinical outcomes. Analyzing the specific influencing factors will be beneficial in guiding clinical decision-making. This article reviews the impact of factors such as advanced age, treatment duration, number of injections, characteristics of neovascular lesions, macular structure, intraocular cytokine levels, and genetics on the response to anti-VEGF therapy. In addition, recent studies have found that pericytes, as cellular components of microvascular walls, can influence the sensitivity to anti-VEGF therapy. This review summarizes the current research on the mechanisms of pericytes in poor or non-response to anti-VEGF therapy and discusses targeted strategies focusing on pericytes.
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SUMMARY: Marein is the main active substance of Coreopsis tinctoria nutt. It not only has anti-oxidation and anti-tumor effects, but also can lower blood lipid, prevent high blood glucose, improve insulin resistance, inhibit gluconeogenesis and promote glycogen synthesis. However, the exact mechanism of its action is still unclear. Here, we explored the effect and mechanism of Marein on insulin resistance. The mice were divided into db/m, db/db, metformin+db/db, and marein+db/db groups. The body weight and kidney weight were recorded. Serum biochemical and renal function tests were measured after 8 weeks of continuous administration. Kidney tissues were subjected to HE staining, PAS staining, and Masson staining. The effect of marein on PI3K/Akt signal and autophagy pathway was detected by Western blot. After 8 weeks of Marein intervention, the body weight and kidney weight of mice did not change significantly, but the fasting blood glucose and blood lipid levels were significantly reduced than db/db group. Marein significantly improved the insulin resistance index, increased serum adiponectin and improved glucose and lipid metabolism disorders of db/db mice. Moreover, marein improved the basement membrane thickness of glomeruli and tubules, improved glomerular sclerosis and tubular fibrosis, as well as renal insufficiency, thereby protecting kidney function and delaying the pathological damage. Furthermore, marein increased the expression of PI3K and the phosphorylation of Akt/Akt (Ser473), and promoted the expression of LC3II/I, Beclin1 and ATG5. Additionally, it promoted the expression of FGFR1 in the kidney of db/db mice, and promoted the increase of serum FGF21 and FGF23. Marein has a protective effect on the kidneys of diabetic mice. It protects diabetic nephropathy by regulating the IRS1/PI3K/Akt signaling pathway to improve insulin resistance. Therefore, marein may be an insulin sensitizer.
RESUMEN: Marein es la principal sustancia activa de Coreopsis tinctoria nutt. No solo tiene efectos antioxidantes y antitumorales, sino que también puede reducir los lípidos en sangre, prevenir la glucemia alta, mejorar la resistencia a la insulina, inhibir la gluconeogénesis y promover la síntesis de glucógeno. Sin embargo, el mecanismo exacto de su acción aún no está claro. Se analizó el efecto y el mecanismo de Marein sobre la resistencia a la insulina. Los ratones se dividieron en grupos db / m, db / db, metformina + db / db y mareína + db / db. Se registró el peso corporal y el peso de los riñones. Se midieron las pruebas de función renal y bioquímica sérica después de 8 semanas de administración continua. Los tejidos renales se sometieron a tinción HE, tinción PAS y tinción Masson. El efecto de la mareína sobre la señal de PI3K / Akt y la vía de autofagia se detectó mediante Western blot. Al término de 8 semanas de tratamiento con mareína, el peso corporal y el peso de los riñones de los ratones no cambiaron significativamente, pero los niveles de glucosa en sangre y lípidos en sangre en ayunas se redujeron significativamente en relación a los del grupo db / db. Marein mejoró significativamente el índice de resistencia a la insulina, aumentó la adiponectina sérica y mejoró los trastornos del metabolismo de la glucosa y los lípidos de los ratones db / db. Además, la mareína mejoró el grosor de la membrana basal de los glomérulos y túbulos, mejoró la esclerosis glomerular y la fibrosis tubular, así como la insuficiencia renal, protegiendo la función renal y retrasando el daño patológico. Además, la mareína aumentó la expresión de PI3K y la fosforilación de Akt / Akt (Ser473), y promovió la expresión de LC3II / I, Beclin1 y ATG5. Además, promovió la expresión de FGFR1 en el riñón de ratones db / db y el aumento de FGF21 y FGF23 en suero. Marein tiene un efecto protector sobre los riñones de ratones diabéticos. Protege la nefropatía diabética regulando la vía de señalización IRS1 / PI3K / Akt para mejorar la resistencia a la insulina. Por tanto, la mareína puede ser un sensibilizador a la insulina.
Subject(s)
Animals , Mice , Insulin Resistance , Chalcones/administration & dosage , Diabetic Nephropathies , Autophagy/drug effects , Blood Glucose , Body Weight/drug effects , Immunohistochemistry , Blotting, Western , Lipids/bloodABSTRACT
Objective:To study the protective effect and mechanism of Ranae Oviductus protein hydrolysate （ROPH） on the expression of pathway-related proteins in ethanol-induced L-02 cell injury. Method:The ROPH was prepared by compound enzymatic hydrolysis. L-02 cell injury model was induced with 400 mmol·L<sup>-1 </sup>ethanol. Cell viability was detected by cell counting kit-8 （CCK-8） assay. Cell cycle and apoptosis were examined by flow cytometry. JC-1/Hochest staining was employed for qualitative investigation. The expression of related proteins in apoptosis， mitogen-activated protein kinase （MAPK） signaling pathway， and pyroptosis in L-02 cells was detected by Western blot. Result:The results of the CCK-8 assay showed that 400 mmol·L<sup>-1 </sup>ethanol could induce L-02 cell injury within 12 hours. Compared with the blank group， the model group showed decreased viability of L-02 cells （<italic>P</italic><0.01）， elevated percentage of the cell cycle in the G<sub>0</sub>/G<sub>1</sub> phase （<italic>P</italic><0.01）， increased total cell apoptosis rate （<italic>P</italic><0.01）， reduced mitochondrial membrane potential （<italic>P</italic><0.01）， up-regulated expression of apoptosis-related proteins ［B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Cytochrome C （Cyt C）， and cysteine-dependent aspartate specific protease-3 （Caspase-3）］ （<italic>P</italic><0.05， <italic>P</italic><0.01） and MAPK signaling pathway-related proteins ［C-Jun amino-terminal kinase （JNK） and p38 MAPK］ （<italic>P</italic><0.05， <italic>P</italic><0.01）， and potentiated expression of pyrolysis-related proteins Caspase-1 and interleukin-1<italic>β </italic>（IL-1<italic>β</italic>） （<italic>P</italic><0.05）. Compared with the model group， the ROPH treatment group exhibited improved cell cycle arrest （<italic>P</italic><0.05， <italic>P</italic><0.01）， diminished total cell apoptosis rate （<italic>P</italic><0.01）， elevated mitochondrial membrane potential in a dose-dependent manner， down-regulated expression of Bax， Cyt C， and Caspase-3 proteins （<italic>P</italic><0.05， <italic>P</italic><0.01）， up-regulated expression of Bcl-2 protein （<italic>P</italic><0.05， <italic>P</italic><0.01）， and a downward trend in expression of proteins related to MAPK signaling pathway and pyrolysis （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:ROPH could inhibit oxidative stress-triggered liver injury in ethanol-induced cells by improving mitochondrial membrane potential， reducing the expression of proteins in the mitochondria-mediated apoptosis pathway， and inhibiting the expression of proteins related to the MAPK signaling pathway and pyrolysis pathway to reduce the mitochondrial dysfunction and inflammatory response in ethanol-induced L-02 liver cells and inhibit oxidative stress， thereby exerting a therapeutic role in alcoholic liver injury.
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Objective: To study the chemical constituents of 95% ethanol aqueous extract of Chuanxiong Rhizoma and the bioactivities of inhibition on nitric oxide (NO) production in lipopolysaccharides (LPS)-activated murine macrophage RAW264. 7 and mouse microglia BV2 cell lines. Methods: The compounds were separated and purified by silica gel column and high performance liquid chromatographies, and their structures were determined by spectroscopic data analysis. Using LPS-activated RAW264. 7 and BV2 cell line models in vitro, all of the isolated compounds were evaluated for the inhibition against NO production. Results: Three butylphthalide derivatives were obtained and identified as Z-3', 8', 3'a, 7'a-tetrahydro-6, 3', 7, 7'a-diligustilide-8'-one (1), Z, Z'-3.3'a, 7. 7'a-diligustilide (2), and chuanliguspirolide (3), respectively. For the inhibition of NO production in the LPS-activated two cell lines, the half inhibitory concentration (IC50) values of compounds 1-3 and indomethacin as a positive control drug in RAW 264. 7 cell line model were (31.60 ± 2.62), (21.20 ± 0.61), (30.12 ± 2.90), and (54.62 ± 7.53) μmol/L, respectively, while IC50 values of compounds 1-3 and curcumin as a positive control drug in BV2 cell line model were (21.99 ± 4.40), (15.43 ± 1.34), (12.20 ± 3.40), and (10.58 ± 1. 41) μmol/L, respectively. Conclusion: Compound 3 named as chuanliguspirolide is a new one. The results of bioactivity assays indicated that compounds 1-3 are potential anti-inflammatory agents.
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In the future,anti-ship missiles(ASM) will be major weapons in the sea war.It is very important to handle the profile of the casualties aboard warships attacked by ASM for development of naval health service.The statistical result shows there is a greater chance of casualties but less chance of warships being sunken by ASM.Besides,medical staffs should pay more attention to the first aid for victims due to blast,burn,smoke inhalation and penetration while the danger of the sea water immersion should not be ignored.
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This paper collected the traditional Chinese medicine (TCM) prescriptions of osteoporosis and osteoarthritis from CNKI to set up database and used TCM inheritance system to analyze the composing principles in prescriptions. We collected 103 and 106 prescriptions for osteoporosis and osteoarthritis respectively, then analyzed inherent principles between herbs in prescriptions by using the TCM inheritance system, and obtained the frequency of each herb, and drug combinations of two diseases' prescriptions. The result showed that the principles in TCM prescriptions of osteoporosis and osteoarthritis are similarities, buch as tonifying kidney and invigorating the circulation of blood, but differences of the specific drugs. That is to say, tonifying kidney is the main method of osteoporosis treatment, however, invigorating the circulation of blood is the main method of osteoarthritis treatment.
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Humans , Blood Circulation , Databases, Factual , Drug Prescriptions , Drugs, Chinese Herbal , Chemistry , Therapeutic Uses , Kidney , Osteoarthritis , Drug Therapy , Osteoporosis , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To understand the comorbid characteristics and distribution of combined treatment of Chinese and Western medicine in depressive patients.</p><p><b>METHOD</b>The descriptive statistic method and association rule were used to analyze the data from 19 general hospitals with 3-A level in China.</p><p><b>RESULT</b>Among the depressive disorder, the most frequent co-morbid physical diseases included hypertension (24.67%), coronary heart disease (16.10%) and cerebral infarction (12.89%), and the proportion of comorbid changes with the increasing age, from 6.51% to 12.55%, 16.33% and 12.47% for hypertension; from 2.79% to 5.69%, 10.17% and 14.22% for coronary heart disease; from 3.72%, 6.27%, 7.70% and 12.25% for cerebral infarction. The use frequency of the antidepressants is 77.18%, and the use frequency of flupentixol & melitracen is 20.95%. The use frequency of Huoxue Huayu Tongluo of traditional Chinese medicine is 59.97%, with that of 27.91% for Ginkgo biloba extract The combined use frequency of Huoxue Huayu Tongluo of TCM and the antidepressants is the highest, especially for the combined use of Shuxuening injection and fluoxetine.</p><p><b>CONCLUSION</b>The most frequent comorbid diseases of depression include three kinds of diseases, such as hypertension, coronary heart disease and cerebral infarction, and its proportion gradually increased with the growth of age. The single use frequency of flupentixol & melitracen and G. biloba extract is the highest, while the combined use of Shuxuening injection and fluoxetine is the highest.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Middle Aged , Young Adult , Anthracenes , Therapeutic Uses , Antidepressive Agents , Therapeutic Uses , China , Combined Modality Therapy , Methods , Depressive Disorder , Drug Therapy , Drug Combinations , Drugs, Chinese Herbal , Therapeutic Uses , Fluoxetine , Therapeutic Uses , Flupenthixol , Therapeutic Uses , Ginkgo biloba , Chemistry , Medicine, Chinese Traditional , Methods , Plant Extracts , Therapeutic UsesABSTRACT
Evaluation of medical assistance capability plays a significant role in improving medical assistance capability and accomplishing a diverse array of military tasks.Based on gray theory and fuzzy mathematics,we put forward a new evaluation model for disaster medical assistance capability,integrating the AHP,the gray cluster analysis and fuzzy evaluation.This model not only reflects the ambiguity of medical assistance capability evaluation but also makes full use of the information,which avoids the flaws caused by single method of fuzzy mathematics.It provides a new way to evaluate disaster medical assistance capability comprehensively and systematically.
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OBJECTIVE: There are some people in population susceptible to acute mountain sickness. Therefore, it is important to analyse, assess and integrate certain research results to build prediction system and mathematics model in order to predict those susceptible people when army goes to tableland.DATA SOURCES: Computer was used to search databases such as Medline, PubMed and PML to find articles regarding prediction of people susceptible to acute mountain sickness from January 1970 to December 2002 with the search words "acute mountain sickness, susceptible population,prediction" . The language was limited to English. At the same time, Chinese Journal Full-text Database, Chinainfo and CBMdisc were searched to find articles from January 1970 to August 2004 by the Chinese language of "acute mountain sickness, susceptible population and prediction". The research targets are susceptible people to acute mountain sickness. At last,trace-back method was used to supplement some literature and monographs.DATA SELECTION: After systematically analyzed and concluded the literature information and screened researches without conducting experiment, full text of the rest literature was searched and used as selective criteria if it could be used as prediction indicator of susceptible people to acute mountain sickness.DATA EXTRACTION: Totally there were 19 prediction indicators generalized. After conducting systemic analysis, Delphi method and analytic hierarchy process(AHP) to the indicators, 13 were selected while the other 6 were excluded.DATA SYNTHESIS: The indicators were classified according to nervous-humoral regulation ability, oxygen capture ability of respiratory system,anti-anoxia ability of central nervous system, mental health and health service ability. Based on these, system analysis, Delphi method and AHP method were used to screen prediction indicators, establish indictor system, confirm the weighted indicators and weighted indicator system as well as standardize the prediction indicators.CONCLUSION: According to the principle of prediction model and model methods, weighted linear synthesis method was used to build the predictive mathematic model. The project of susceptible population prediction of acute mountain sickness in army when entering tableland urgently was proposed combining the health service ability of army.
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Objective To discuss the influence of new weapons in the high-tech war on eye wounds in order to develop the new time-effect treatment strategy. Methods The data of wounds in recent wars were investigated, the causes of eye wounds in high-tech war were analyzed, and the new concept of time effect treatment strategy for eye wounds was proposed. Results The weapons in high-tech war have developed into mass destruction and precisely guided munitions. The main cause of eye wounds is the shrapnel and some new wounds will occur such as eye wounds caused by laser or chemical toxin, radiation cataract. Conclusion In the future high-tech war, the major type of eye-wounds may still be the explosion injury and some new types of eye wounds should be paid more attention. The time-effect treatment strategy should be implemented to meet the demand of improving eye wound therapy and promoting the development of ophthalmology.