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The mammalian epididymis not only plays a fundamental role in the maturation of spermatozoa, but also provides protection against various stressors. The foremost among these is the threat posed by oxidative stress, which arises from an imbalance in reactive oxygen species and can elicit damage to cellular lipids, proteins, and nucleic acids. In mice, the risk of oxidative damage to spermatozoa is mitigated through the expression and secretion of glutathione peroxidase 5 (GPX5) as a major luminal scavenger in the proximal caput epididymidal segment. Accordingly, the loss of GPX5-mediated protection leads to impaired DNA integrity in the spermatozoa of aged Gpx5
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Objective@#To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters.@*Methods@#By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used.@*Results@#The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889.@*Conclusions@#The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction.
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Objective:To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters. Methods:By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used. Results:The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889. Conclusions:The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction.
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BACKGROUND: Superparamagnetic iron oxide (SPIO) labeling technology is a classic noninvasive tracing method, which has been widely used in the stem cell transplantation. Induced pluripotent stem cells (iPSCs) are currently one of the most promising seed cells for cell transplantation. Whether SPIO labeling can also be used to noninvasively trace induced pluripotent stem cells is rarely reported, and concern has been raised about whether SPIO markedly impacts the differentiation of iPSCs. OBJECTIVE:To investigate the effects of SPIO labeling on the differentiation of iPSCs in vitro. METHODS: Rat fibroblasts were isolated and cultured. Efficient recombinant vector and plasmids that were packaged by virus and contained target genes (Oct4, Sox2, Klf4 and c-Myc) were transfected into 293T cells for virus packaging and production. The packaging lentiviral vectors that contained target genes infected rat fibroblasts to obtain iPSCs. SPIO-labeled (experimental) or unlabeled (control) iPSCs were subjected to neural induction and differentiation. Prussian blue staining and transmission electron microscope observation were performed for SPIO-labeled iPSCs. Immunohistochemical method was used to detect neuron-specific enolase expression after induced differentiation. Flow cytometry was used to detect the proportion of neurons and glial cells differentiated from iPSCs. RESULTS AND CONCLUSION: There were dense iron particles in the cytoplasm of SPIO-labeled iPSCs shown by Prussian staining and under transmission electron microscope. Differentiated iPSCs were positive for neuron-specific enolase. In addition, the proportion of neurons and glial cells showed no difference between the experimental and control groups. To conclude, SPIO labeling has no obvious effect on the capacity of iPSCs differentiating into neurons. Reasonable application of this new cell labeling technique will promote the development of seed cells in regenerative medicine.
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AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.
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Objective To research the effect and safety of Reteplase for Injection combined with reduced glutathione in the treatment of acute ST segment elevation myocardial infarction.Methods Patients with acute ST segment elevation myocardial infarction (90 cases) in Jianli People's Hospital from February 2015 to February 2016 were selected and divided equally into A,B and C groups according to different treatment methods.Totally 30 patients in group A were given conventional therapy,30 patients in group B were given reduced glutathione for treatment on the basis of conventional treatment,and the other 30 in group C were given combined therapy of reteplase and reduced glutathione on the basis of conventional treatment.The vascular recanalization rate,improvement of effective indicators including creatine kinase isoenzyme (CKMB),troponin Ⅰ (cTnⅠ),left ventricular end diastolic diameter (LVEDd) and left ventricular ejection fraction (LVEF),oxidative stress kinase including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px),and incidence of adverse events of patients were compared among the three groups.Results After thrombolysis,the vascular recanalization rate of group C at different time points (2 h,6 h and 12 h) showed significant difference compared with those of groups A and B (P ＜ 0.05).After the treatment,the effective indicators of the three groups were both significantly improved (P ＜ 0.05),and group C improved more significantly than groups A and B (P ＜ 0.05).After the treatment,the SOD and GSH-Px of groups B and C both significantly improved than group A (P ＜ 0.05),but there was no significant difference between groups B and C.After treatment,the incidence of adverse events of group C was significantly lower than those of groups A and B (P ＜ 0.05).Conclusion Reteplase for Injection combined with reduced glutathione has significant curative effect in the treatment of acute ST segment elevation myocardial infarction,which can effectively improve the cardiac function and inhibition of oxidative stress.It is of higher security but with lower incidence of adverse events.
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<p><b>BACKGROUND</b>Talaromyces (Penicillium) marneffei (TM) is an emerging dimorphic human pathogenic fungus that is endemic to Southeast Asia. TM mostly occurs as an opportunistic infection in patients with human immunodeficiency virus (HIV). The objective of this study was to compare the clinical and laboratory parameters of patients with TM infections who were HIV-positive and HIV-negative and to assess therapies and outcomes.</p><p><b>METHODS</b>This was a retrospective analysis of 26 patients diagnosed with disseminated TM infection from September 2005 to April 2014 at Fujian Provincial Hospital, China.</p><p><b>RESULTS</b>Patients with TM infection tend to present with fever, weight loss, and anemia. The time from symptom onset to confirmed diagnosis was greater for HIV-negative patients (n = 7; median: 60 days, range: 14-365 days) than for HIV-positive patients (n = 19; median: 30 days, range: 3-90 days, Mann-Whitney U = 31.50, P= 0.041). HIV-negative patients were more likely to have dyspnea (57.1% vs. 5.3%, χ2 = 8.86, P= 0.010), low neutrophil count (Mann-Whitney U = 27.00, P= 0.029), high CD4 count (Mann-Whitney U = 0.00, P= 0.009), and high lymphocyte count (Mann-Whitney U = 21.00, P= 0.009). There were no significant differences in other demographic, clinical, or biochemical characteristics. Among all the patients, 12 HIV-positive patient and 1 HIV-negative patient received amphotericin and fluconazole treatment, 9 of whom improved, 1 died, 2 had kidney damage, 1 had hypokalemia due to exceeded doses.</p><p><b>CONCLUSIONS</b>HIV-negative patients with TM infections tend to have a longer diagnostic interval, a higher percentage of dyspnea, higher levels of CD4 and lymphocytes, and lower neutrophil counts than TM infection in HIV-positive patients. Treatment programs with amphotericin and fluconazole are mostly effective.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Recuento de Linfocito CD4 , Infecciones por VIH , Micosis , Diagnóstico , Quimioterapia , Alergia e Inmunología , Estudios Retrospectivos , TalaromycesRESUMEN
Resveratrol is a natural polyphenolic substance with a variety of bioactivities,such as anti-oxidation,anti-tumor and apoptosis induction.This paper summarized the recent research progress of resveratrol induced cancerous cell apoptosis,including human cervical cancer HeLa cells,leukemia cells,hepatoma cells and gastric cancer cells,and analyzed the molecular mechanism behind resveratrol-induced cell apoptosis in tumor.In addition,resveratrol was investigated by blocking the cell cycle,regulating the expressions of related genes and proteins and mitochondria-induced apoptosis pathways as a potential application of cancer drugs to clinical researches.
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AIM: To study the changes of brain - derived neurotrophic factor ( BDNF ) expression in gene modified bone marrow mesenchymal stem cells ( BMSC) . ●METHODS:BMSC were divided into blank control group ( without transfected BMSC ) , negative control group ( empty vector without BDNF gene transfected BMSC) and experimental group ( BDNF gene transfected BMSC) . The expression of BDNF mRNA in BMSC was measured by Realtime PCR, and the expression of BDNF in BMSC was measured by ELlSA. ●RESULTS:The BDNF mRNA expressions of 3, 4, 5, 6, 7 and 8-generation BMSC cells in the experimental group were higher than those in the blank control group and negative control group. The differences were statistically significant (P3: F=491. 788, P ●CONCLUSION:Long-term expression of BDNF in BMSC can be enhanced by genetic engineering.
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Abstract?AIM: To investigate the influence on blood-retina barrier after intense light exposure in rats.?METHODS: The rats were randomly divided into light exposure group and control group. Rats in light exposure group were exposed in white light (10000lux, 12h on-off, continuing 1-14d) .Rats in control group were only exposed in natural light.The eyes of the rats in the two groups were removed when the rats in light exposure group acceptted intense light after 1, 3, 7 and 14d.We observed the change of retinal structure using hematoxylin-eosin ( HE ) staining, and observed the change of retinal ultrastructure using electron microscope.We quantified the change of retinal vascular permeability using laser scanning confocal fluorescence microscope and spectrophotometry after perfusion of Evans-blue, to evaluate the change of blood-retinal barrier.?RESULTS: At 1d after intense light exposure, the retinal ultrastructure of rats changed, such as denaturation of photoreceptor cells and falling of membranous disc outer segment and thinning of the outer nuclear layer thickness, and so on;and the longer the rats exposure to intense light, the more serious change of the retinal ultrastructure were found.At 3d later, photoreceptor cells began apoptosis.At 14d later, the outer nuclear layer became thinner obviously, and the number of cells reduce obviously.At 1d after intense light exposure, EB leaked from the retinal vascular, and at 14d later the leaking of EB was more obvious.?CONCLUSION: The photoreceptor cell of the outer nuclear layer of retina will degenerate and apoptosis, and the outer nuclear layer will be thinner, and the structure and function of blood-retinal barrier will be destroied, if the eyes of rats exposed in intense light.
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In this paper, a qualitative and quantitative analysis method was established for the scopoletin in herba Artemisiae hedinii hy HPLC. The chromatographic column was Agilent TC-C18 (4.6 mm×250 mm, 5μm);the tem-perature of column was 30℃, eluted with a mobile phase consisted of methanol and 0.3%phosphoric acid solution and gradient elution at a flow rate of 1.0 mL·min-1. The detection wavelength was 344 nm. The results showed that the mass concentration and peak area of scopoletin had a good linear relationship within the range of measurement (r=0.999 9). The reproducibility was good;and the RSD was 1.51%. The average sample recovery rate was 101.42%. It was concluded that the established method was stable, reliable and simple. It provided theoretical evidences for the quality evaluation and quality control of herba A. hedinii.
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In order to optimize extraction process conditions of tannins from Geranium orientali-tibeticum by supercritical CO2, the content of tannins was determined by phosphomolybdium tungsten acid-casein reaction, with extraction pressure, extraction temper- ature and extraction time as factors, the content of tannins from extract of G. orientali-tibeticum as index, technology conditions were optimized by orthogonal test. Optimum technology conditions were as follows: extraction pressure was 25 MPa, extraction temperature was 50 °C, extracted 1.5 h. The content of tannins in extract was 12.91 mg x g(-1), extract rate was 3.67%. The method established could be used for assay the contents of tannin in G. orientali-tibeticum. The circulated extraction was an effective extraction process that was stable and feasible, and that provides a way of the extraction process conditions of tannin from G. orientali-tibeticum.
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Dióxido de Carbono , Química , Fraccionamiento Químico , Métodos , Medicamentos Herbarios Chinos , Química , Geranium , Química , TaninosRESUMEN
OBJECTIVES@#To investigate the function of cytokines, chemokines, and regulatory T cells (Tregs) in the pathogenesis of type 1 diabetes mellitus (T1DM) in children.@*METHODS@#A total of 35 children with T1DM and 30 healthy controls were enrolled in this study. Levels of serum cytokines (IL-1α, IL-6, IL-10, IL-12, and TNF-α) and chemokines (MIP-1α, MIP-1α and MCP-1) were detected by enzyme-linked immunosorbent assay. Peripheral blood mononuclear cells (PBMCs) were isolated and culture supernatant of phytohaemagglutinin (PHA)-stimulated PBMCs was subjected to ELISA for levels of cytokines (IL-1α, IL-6, IL-10, IL-12 and TNF-α) in T1DM and control group. Furthermore, flow cytometry was used to determine the percentage of Tregs in PBMCs of two groups.@*RESULTS@#Levels of serum cytokines including IL-1α, IL-6, IL-10 and TNF-α as well as chemokines, such as MIP-1α and MIP-1α in children with T1DM children were significantly higher than those in healthy controls (P<0.05, respectively). PBMCs with PHA stimulation in T1DM group secreted more IL-1α and TNF-α (P<0.05, respectively), but less IL-10 (P<0.05), as compared with control group. Furthermore, the proportion of CD4(+), CD25(+), Foxp3(+), Tregs in PBMCs isolated from children with T1DM was obviously lower than those in healthy controls (P<0.05).@*CONCLUSIONS@#Immune dysfunction, with upregulation of inflammatory factors such as IL-1α, IL-6, TNF-α and MIP-1α, downregulation of IL-10 and Tregs, plays an important role in the pathogenesis of T1DM in children.
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Objective To demonstrate the feasibility, effectiveness and reliability of combination of procedure for prolapse and hemorrhoids (PPH) and excision-closed hemorrhoidectomy (ECH) for severe hemorrhoids. Methods Sixty patients with Ⅲ- Ⅳ degree hemorrhoids which left remnant prolapsed hemorrhoids after stapled hemorrhoidopexy, were randomly divided into group A (PPH with additional ECH group,30 cases.) and group B [PPH with additional Milligan-Morgan hemorrhoidectomy (MMH) group, 30 cases]. Ten indexes of the results in two groups were compared. ResultsThe hemorrhoidectomy lasted (35.6 ± 5.3) min in group A and (47.3 ± 10.2) min in group B. The indexes related to 24 h visual analogue scale (VAS) after operation were (2.9 ± 1.2) scores in group A and (5.7 ± 1.9) scores in group B. Wound healing time were (6.3 ± 1.4) d in group A and (28.5 ± 4.8) d in group B. Many indexes showed the results in group A was better than those in group B ( P ＜ 0.05 ). The six-month follow-up visits after operation showed no complications, such as recurrence in either group. ConclusionsCombination treatment for Ⅲ -Ⅳ degree hemorrhoids with PPH with additional ECH is a easy,safe,reliable,minimal-aching and quick-recovering method of hemorrhoidectomy with favorable results than PPH with additional MMH.
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<p><b>OBJECTIVE</b>To establish a mice model with selective inactivation adenosine A2A receptors (A2ARs) in peripheral white blood cells (PWBC).</p><p><b>METHODS</b>A2ARs were selectively inactivated in PWBCs by transplanting bone marrow cells (BMCs) from A2AR knockout (KO) mice into their wild type (WT) littermates after a single total body irradiation of 9.5 Gy or fractionated total body irradiation of 6.2 Gy x 2. The efficiency of reconstitution of bone marrow-derived cells in chimeric mice was assessed.</p><p><b>RESULTS</b>PCR band patterns changed from the recipient pattern (one band of 330 bp) to the donor (two bands of 300 and 330 bp) pattern. Immunohistochemistry analysis showed that 10.21% of cells were A2AR+ in PWBCs in KO--> WT mice, whereas 96.72% of cells were A2AR+ in WT mice. The survival rates of mice irradiated with 6.2 Gy x 2 and transplanted with more than 6 x 10(6) BMCs were about 91%.</p><p><b>CONCLUSION</b>A murine model of selective inactivation adenosine A2A receptors in PWBCs was established successfully.</p>
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Animales , Ratones , Eliminación de Gen , Ratones Endogámicos C57BL , Ratones Noqueados , Modelos Animales , Receptor de Adenosina A2A , GenéticaRESUMEN
Objective To evaluate the effect of preoperative radiotherapy and postoperative brachy radiotherapy (POBRT) on patients with primary liver cancer(PLC). Methods 50 patients with PLC were randomly divided into 2 groups:(1)Radiotherapy group, 25 patients who underwent preoperative radiotherapy 14～17 days before hepatectomy and POBRT 3～10 days after hepatectomy ;(2)Control group,25 cases who underwent hepatectomy only. In radiotherapy group, before heptectomy, the single-dose 6Gy per time and 3 sessions were given to each patient. 3～6 ductus were placed for POBRT during operation,and 10Gy of POBRT per time and 2～4 sessions were given postoperatively. In control group,no radio cherapy was given before and after hepatectomy. Results In radiotherapy group, the cancer shrank significantly after preoperative radiotherapy (P
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Objective To improve the CR image quality and to promote the digital image standard constitution by analyzing the common problems and malpractiees in application of computed radiography. Methods Phenomenon and reasons of 107 CR junk-films from nine three-"A"-hospitals were analyzed, discussed,recorded,and statistised by 20 radiologists,radiographers and engineers.Results Among 107 junk fihns,there are 36 cases(33.64%)of incorrect operations,29 cases(27.10%)of artifacts in reading and transferring the data of IP,15 cases(14.02%)of artifacts in IP system,and 13 cases (12.15%)of selection of inappropriate radiographic parameters,and 9 eases(8.41%)of printer-failures, and 5 eases(4.67%)of inappropriate post-processing techniques.By analyzing the reasons of 107 junk films we found that 60.74% were due to less responsibilities and incorrect operations,and 35.51% were due to new problems in CR techniques,and other were due to inappropriate post-processing techniques. Conclusion Responsibilities,operation regulations,digital image quality standards,studying of new techniques and appropriate use of the post-processing techniques are the key points for improving the CR image quality and the diagnosis level.
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0.05). Conclusion The oral sodium phosphate preparation is superior to magnesium sulfate before colorectal surgery. Single-day preoperative bowel preparation with oral sodium phosphate is suitable for elective colorectal surgery.