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Accumulating evidence has shown that the cell-penetrating peptide TAT can be applied to deliver different types of drug molecules, including nucleic acids, proteins and small molecule drugs. Usually TAT delivers cargoes on the basis of their covalent bonds or non-covalent interactions. However, there are few reports on the delivery of proteins by TAT in a non-covalent manner, and no quantitative comparisons have been made on the protein delivery ability of TAT in fusion and non-fusion manners. In order to explore the ability of TAT to deliver proteins in non-fusion manner, here we used fluorescence microscopy and flow cytometry to investigate the ability of TAT to deliver enhanced green fluorescent protein (EGFP) into non-small cell lung cancer cells A549 in a non-fusion manner. It was found that TAT could deliver EGFP into A549 cells, and its delivery ability was positively correlated with its concentration. In addition, the fusion protein TAT-EGFP was overexpressed and purified, and its permeability across cell membrane was also investigated. In this paper, based on quantitative comparison, we found that the delivery of EGFP by TAT in fusion manner is significantly efficient than that of TAT in non-fusion manner. This is the report that TAT can deliver EGFP in a non-fusion manner. Although its delivery efficiency remains to be improved as compared with the fusion manner, the non-fusion manner has shown incomparable advantages in ease of operation, suggesting that it is also a candidate for delivery strategy in the future.
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Objective:To observe the effect of modified Wuzi Yanzongwan periodic staging treatment on the outcome of assisted pregnancy in patients with poor ovarian response （POR） and kidney deficiency syndrome. Method:One hundred and four patients were randomly divided into observation group and control group， with 52 cases in each group. Both groups received gonadotropin releasing hormone （GnRH） antagonist regimen. The patients in control group additionally took Bushen Yutaiwan orally， 5 g/time， 3 times/day. The patients in observation group additionally took modified Wuzi Yanzongwan during pre-ovulation and post-ovulation periods， 1 dose/day. The treatment courses were 3 menstrual cycles （or termination after clinical pregnancy） in both groups. The number of eggs obtained， the number of available embryos， the number of fertilization， the number of high-quality embryos， the number of embryos implanted， the number of cycles cancelled， and the clinical pregnancy were recorded. Human chorionic gonadotropin （HCG） was injected， and then follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， estrogen （E<sub>2</sub>）， anti-Müllerian hormone （AMH）， basal antral follicle count （AFC） and endometrial thickness were measured daily. The number of days and dosage of Gn used， scores of kidney deficiency syndrome were recorded before and after treatment， and the adverse reactions during the study period were recorded. Result:The number of eggs captured， rate of harvested eggs， number of available embryos， rate of available embryos， number of high-quality embryos， rate of high-quality embryos， and fertilization rate in observation group were higher than those in control group （<italic>P</italic><0.05 or <italic>P</italic><0.01）. The cycle cancellation rate was lower than that in the control group； the embryo implantation rate and clinical pregnancy rate were superior than those in control group， but the difference was not statistically significant. The FSH level and FSH/LH ratio in observation group were lower than those in control group during HCG day （<italic>P</italic><0.01）， while E<sub>2</sub>， AMH， AFC and endometrial thickness were higher than those in control group （<italic>P</italic><0.01）. Simultaneously， the number of days and amount of Gn used in observation group was lower than that in control group （<italic>P</italic><0.01）. Conclusion:The Modified Wuzi Yanzongwan periodic staging treatment combined with GnRH antagonist scheme for patients with POR kidney deficiency syndrome， can regulate the level of endocrine hormones， promote follicular development， improve ovarian reserve， increase the number of eggs obtained， improve egg quality， help improve pregnancy outcomes， and increase the chances of successful pregnancy with assisted reproductive technology. It is worthy of further clinical research.
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OBJECTIVE@#To analyze the changes and characteristics of pediatric outpatient visits in a general hospital before and after the coronavirus disease (COVID-19) epidemic.@*METHODS@#Based on the registration data of pediatric outpatient visits in the information system (HIS)of Beijing Tsinghua Changgung Hospital, from January 1 2018 to December 31 2020, aged 0 to 16 years, we analyzed the changes of outpatient visits before and after the epidemic, focusing on respiratory infection including influenza. The relationship between the outpatient visits and age and quarterly distribution were also studied.@*RESULTS@#(1) Respiratory infection accounted for the majority of outpatient visits in 2018 and 2019 (60.6% and 60.5%, respectively). Non-respiratory infection accounted for the main proportion of outpatient visits in 2020, while respiratory infection accounted for only 47.4%. Annual respiratory infection visits, respiratory infectious diseases visits especially influenza visits all decreased significantly in 2020 compared with that in 2018 and 2019 (P < 0.05). (2)Respiratory infection visits were highest in the infant group, lowest in the school age group (P < 0.05) and highest in the fourth quarter each year. It decreased significantly in the second quarter of 2020 with statistical significance when compared with the other quarters of 2020(P < 0.05). (3)Influenza accounted for the highest proportion of respiratory infectious diseases visits in each year. It was highest in first quarter, which was significantly different from the other quarters of the year (P < 0.05). There were different distributions of influenza visits throughout 2018 and 2019, while it was only distributed in the first quarter and 99% in January in 2020.@*CONCLUSION@#The respiratory infection and influenza visits have decreased significantly in our pediatric outpatient department after the COVID-19 epidemic, which is considered closely related to the lifestyle and personal protection after the epidemic. It is recommended that health education on respiratory infection and influenza prevention should be strengthened, especially in winter and spring, to promote the development of good respiratory and hand hygiene habits.
Subject(s)
Child , Humans , Infant , COVID-19 , Hospitals, General , Influenza, Human/epidemiology , Outpatients , Pandemics , SARS-CoV-2ABSTRACT
OBJECTIVE: To study the effect of serum containing Jieduquyuziyin-prescription (JP) on signal pathway of interleukin-1 receptor-associated kinase 1 (IRAK1) in mononuclear macrophages of mice stimulated by lipopolysaccharide (LPS), and to explore the effect of Jieduquyuziyin-prescription on IRAK1 and NF-κB inflammatory signaling pathways, which providing a good theoretical support for its anti-inflammatory clinical medication. METHODS: In this study, mice mononuclear macrophages cultured in vitro were randomly divided into blank group, LPS group, JP serum group, blank serum group, LPS plus JP serum group, LPS plus blank serum group, IRAK1 inhibitor group, inhibitor plus LPS group, inhibitor plus JP serum group and inhibitor plus blank serum group. After intervention for 24 h, the activity of JP on macrophages was tested by CCK8 method. The IRAK1 expression in macrophages was tested by immunofluorescence chemical staining. The content of TNF-α in the supernatant of the cells was detected by ELISA. The mRNA expressions of IRAK1, NF-κB, TNF-α and IL-6 were detected by RT-PCR. The protein expressions of IRAK1, p-IRAK1 and NF-κB were detected by Western-blot. The LC-MS was used to detect the active ingredients in JP serum. RESULTS: The results show that 2.5% of JP serum is the optimal concentration. Jieduquyuziyin-prescription could down-regulate the expression of TNF-α and IL-6 and inhibit the expression of IRAK1 and activate NF-κB(P<0.05). Paeoniflorin and ferulic acid were detected in the JP serum. CONCLUSION: Jieduquyuziyin-prescription can inhibit the expression of IRAK1 and NF-κB in mouse monocyte-macrophage cells after LPS stimulation and provide a good theoretical support for its anti-inflammatory clinical medication.
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OBJECTIVE@#To investigate the incidence of congenital anomalies of the kidney and urinary tract (CAKUT) in neonates, and to evaluate the value of urinary ultrasound screening in the early postnatal period.@*METHODS@#The neonates born or treated in Beijing Tsinghua Changgung Hospital affiliated to Tsinghua University between January 2016 and December 2018 accepted the urinary ultrasound screening, and the neonates with problem were followed up. In the meanwhile, the maternal pregnancy data were analyzed to screen out the risk factors associated with the onset of CAKUT.@*RESULTS@#(1)A total of 2 655 neonates were screened by ultrasonography, of whom 82 neonates had been diagnosed with CAKUT (male: 60 cases, female: 22 cases), the positive rate was 3.1% (82/2 655). There were 66 cases of hydronephrosis, 6 cases of duplicate kidney, 2 cases of multiple renal cysts, 2 cases of renal cystic dysplasia, 1 case of medullary sponge kidney, 3 cases of small kidney, 1 case of isolated kidney, and 1 case of horseshoe kidney. (2)Of the 66 children with hydronephrosis, 4 cases were lost to the follow-up; 8 cases were followed for less than six months with no significant changes found, and still in the follow-up observation; 54 cases were followed up for 1 year, among which 32 cases were returned to normal within 1 year, 3 cases were alleviated, 7 cases were aggravated, and 12 cases were unchanged. One case underwent surgery for repeated urinary tract infections and decreased renal function. (3) Abnormal fetal urinary ultrasound in the late pregnancy was found to be the most common in the high risk factors of CAKUT. There were 44 high-risk newborns with abnormal fetal urinary ultrasound, and 35 cases of CAKUT were diagnosed after birth. The incidence rate was 79.5%(35/44). (4)Among the 2 655 newborns screened, 2 611 newborns had normal antenatal urinary ultrasonography. Among these neonates with normal urinary ultrasound during pregnancy,47 cases of CAKUT were diagnosed after birth, with an incidence of 1.8% (47/2 611).@*CONCLUSION@#The most common CAKUT in neonates is hydronephrosis and most cases with hydronephrosis had a good prognosis, but they should be followed up regularly. Urinary ultrasound screening for neonates, especially those high-risk neonates with abnormal fetal urinary ultrasound, has important clinical implications for the early detection of CAKUT.
Subject(s)
Child , Female , Humans , Infant, Newborn , Male , Pregnancy , Follow-Up Studies , Hydronephrosis , Kidney , Ultrasonography , Urinary TractABSTRACT
Objective To study the effects of Jiedu Quyu Ziyin Prescription (JQZP)-treated freeze dried powder and drug-containing serum on the inflammatory signal pathway of monocyte-macrophage induced by LPS (lipopolysaccharide) in mice. Methods Monocyte-macrophage cells were cultured in vitro and randomly divided into blank group, LPS stimulation group, drug-containing serum group, freeze dried powder group, LPS + drug-containing serum group, and LPS + freeze dried powder group. After 24 h intervention, the optimal concentrations of drug-containing serum and freeze dried powder were screened by CCK8 method and the cell viability was measured respectively. The content of tumor necrosis factor (TNF-α) in cell serum was measured by ELISA. Real-time PCR was employed to test the expression of TNF-α mRNA and nuclear transcription factor kappa-light-chain-enhancer of activated B cells (NF-κB). Western-blot was used to detect the expression of NF-κB protein. The LC-MS was used to detect the active ingredients in the drug-containing serum. Results Compared with the blank group, the expression of TNF-α level, NF-κB, TNF-α mRNA and NF-κB protein in LPS stimulation group were significantly increased (P < 0.05). Compared with the LPS stimulation group, the TNF-α level, NF-κB, TNF-α mRNA and the expression of NF-κB protein in the LPS plus serum group were significantly lower than those in the LPS plus freeze-dried powder group (P < 0.05). Paeoniflorin and ferulic acid were detected in the drug-containing serum. Conclusion JQZP freeze-dried powder and drug-containing serum all have the effect of inhibiting the inflammatory signaling pathway.
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This study is to establish the UPLC fingerprint of red ginseng. The separation was performed on a Waters Acquity BEH C₁₈ column (2.1 mm × 50 mm,1.7 μm), with the mobile phase consisting of acetonitrile and water for gradient elution. The detection wavelength was set at 203 nm. The UPLC fingerprint of red ginseng was established by using sample chromatography of 22 different purchase areas and 26 common peaks were found. Compared with the reference substances, 11 of the common peaks were identified as ginsenosides Rg₁, ginsenoside Re, ginsenoside Rf, ginsenoside Rh₁, ginsenoside Rg₂, ginsenoside Rb₁, 20(S)-ginsenoside F₁, ginsenoside Rb₂, ginsenoside Rb3, 20(S)-ginsenoside Rg₃ and 20(R)-ginsenoside Rg₃, respectively. It is worth noting that 20(S)-ginsenoside Rg₃ and 20(R)-ginsenoside Rg₃ are the characteristic ingredients of red ginseng, and they could be used not only for distinguishing red ginseng and ginseng, but also for process controlling of the preparation of red ginseng. The similarity was analyzed with' Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica, and the similarity of 18 batches samples is up to 0.9. Compared to the literature methods, the method is simple, time-saving,specific for the separation of ginsenosides from red ginseng. So, this method could be used for the species identification and quality control of ginseng, red ginseng and American ginseng, and it will alsoprovide a theoretical basis of raising quality standards of the above mentioned Chinese herb medicines.
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Using reversed-phase high performance liquid chromatography, nine ginsenosides were simultaneously separated on an UltimateC₁₈ column with high-resolution and high purity of each chromatographic peak. Adopting the QAMS quality evaluation model for traditional Chinese medicines, ginsenoside Rb₁ was used as the internal reference substance, and the relative correction factors (RCFs) and the relative retention values (RTRs) of ginsenosides Rg₁, Re, Rf, Rb₁, Rc, Rb₂, Rb₃, Rd and 20 (S)-ginsenoside Rg₃ to ginsenoside Rb₁ were calculated individually. Through a series of methodology evaluations, and positioned by the red ginseng reference chromatograph and RTVs, nine ginsenosides in red ginseng were simultaneously assayed only by quantitative determined ginsenoside Rb₁.
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<p><b>OBJECTIVE</b>To study the efficacy of Huai Qi Huang granules in the treatment of childhood primary nephrotic syndrome.</p><p><b>METHODS</b>Between July 2009 and December 2011, patients who were admitted and diagnosed for the first time as childhood primary nephrotic syndrome were randomized into a treatment group (Huai Qi Huang granules plus glucocorticoid; n=23) and a control group (glucocorticoid alone; n=19) for a prospective study. The two groups were compared for regression time of edema, time to urinary protein clearance, relapse rate, incidence of infection, dosage of glucocorticoid, and humoral and cellular immunological indicators.</p><p><b>RESULTS</b>There were no significant differences in regression time of edema, time to urinary protein clearance, and relapse rate between the treatment and control groups (P>0.05). The treatment group had significantly lower incidence of infection and daily dose of glucocorticoid (at month 6) than the control group (P<0.05). Humoral and cellular immunological indicators showed no significant differences between the two groups (P>0.05). No Huai Qi Huang-related adverse events were observed in this study.</p><p><b>CONCLUSIONS</b>Huai Qi Huang granules treatment can reduce the dose of glucocorticoid and the incidence of infection in children with primary nephrotic syndrome and has a favourable safety.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Astragalus propinquus , Drugs, Chinese Herbal , Therapeutic Uses , Glucocorticoids , Therapeutic Uses , Nephrotic Syndrome , Drug Therapy , Prospective StudiesABSTRACT
<p><b>OBJECTIVE</b>To establish an HPLC-DAD method for simultaneous determination of eight ginsenosides in Renshenshouwu capsules.</p><p><b>METHOD</b>Ultimate C18 column (4.6 mm x 250 mm, 5 microm) was adopted for gradient elution, with acetonitrile and water as the mobile phase. The flow rate was 1 mL x min(-1), the column temperature was set at 30 degrees C, and the detection wavelength was 203 nm.</p><p><b>RESULT</b>A good linearity was observed in the range of 0.242-12.1, 0.222-11.1, 0.251-25.1, 0. 245-24.5, 0.232-23.2, 0.232-23.2, 0.264-26.4, 0.244-24.4 microg for ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd and 20(S)-ginsenoside Rg3, respectively, with the average recoveries of 102.7%, 103.2%, 101.6%, 101.2%, 102.0%, 100.7%, 101.9%, 102.0%, respectively.</p><p><b>CONCLUSION</b>The method was so simple, accurate and effective that it could be used for quality control of the above eight components in Renshenshouwu capsules.</p>
Subject(s)
Capsules , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , GinsenosidesABSTRACT
OBJECTIVE: To develop an HPLC-ELSD method for simultaneous determination of the contents of eight ginsenosides in Renshenshouwu capsules (traditional Chinese medicines). METHODS: The analysis was performed on an Lltimate C18 column (4.6 mm × 250 mm, 5 μm) eluted with mobile phase consisting of acetonitrile and water in gradient mode at a flow rate of 1.0 mL · min-1. The column temperature was set at 30°C. The ELSD drift tube temperature was set at 100°C, and the gas flow rate was set at 2.5 L · min-1. RESULTS: Good linearity(r > 0.9995) was obtained in the range of 0.483-12.1 μg, 0.444-11.1 μg, 0.502-12.6 μg, 0.490-12.3 μg, 0.455 23.2 μg, 0.464-11.6 μg, 0.264-13.2 μg and 0.244-12.2 μg for ginsenoside Rg1, ginsenoside Re, gin-senoside Rb1, ginsenoside Rc, ginsenoside Rb2, ginsenoside Rb3, ginsenoside Rd and 20(S)-ginsenoside Rg3, respectively. The average recoveries of the eight compounds were 101.9%, 104.5%, 98.3%, 97.9%, 97.8%, 101.2%, 100.6% and 99.0% Respectively. CONCLUSION: The method is reliable, simple, precise, and can be used for the quality control of Renshenshouwu capsules.
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<p><b>OBJECTIVE</b>To explore the role of autophagy in quercetin (Que)-induced apoptosis in human bladder carcinoma BIU-87 cells in vitro.</p><p><b>METHODS</b>To determine the proliferative inhibition by MTT colorimetric assay after treating BIU-87 cells with quercetin at various concentrations. To identify autophagy and apoptosis in the BIU-87 cells after Que treatment by monodansylcadaverin (MDC) and Hoechst 33258 fluorescent staining, respectively. To examine the cytotoxic effect of Que and influence of autophagy on apoptosis by studying LDH leakage rate and flow cytometry, after blocking the autophagy with 3-methlyadenine (3-MA), a specific autophagy inhibitor.</p><p><b>RESULTS</b>There was an obvious inhibitory effect of Que on the proliferation of BIU-87 cells in a time- and dose-dependent manner. The inhibition rate of BIU-87 cells after 200 µmol/L Que treatment for 72 hours was 89.2%. Autophagy and apoptosis were induced and detected in Que-treated BIU-87 cells and autophagy occurred earlier than apoptosis. The apoptosis peak became much higher after the autophagy was blocked. Whenever the autophagy was blocked before or after Que treatment, the Que-induced cytotoxicity in BIU-87 cells was enhanced.</p><p><b>CONCLUSIONS</b>Quercetin significantly inhibits the proliferation of BIU-87 cells, and the autophagy is induced earlier than apoptosis. In the process of Que-induced apoptosis of BIU-87 cells, autophagy may play a protective role at the initiation phase, delay apoptosis and reduce the Que-induced death of BIU-87 cells.</p>
Subject(s)
Humans , Adenine , Pharmacology , Antioxidants , Pharmacology , Apoptosis , Autophagy , Physiology , Cell Line, Tumor , Cell Proliferation , Dose-Response Relationship, Drug , L-Lactate Dehydrogenase , Metabolism , Quercetin , Pharmacology , Urinary Bladder Neoplasms , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of different chromatographic conditions on QAMS relative correlation factors (RCF) and relative retention values (RT(R)).</p><p><b>METHOD</b>C18 columns were used with methyl alcohol-0.4% phosphoric acid water (85: 15) as the mobile phase. The detection wavelength was 254 nm, the column temperature was 30 degrees C, and the flow rate was 1.0 mL x min(-1). The five anthraquinones in Rhei Radix et Rhizoma were selected to be the objects of study. The RCF and RT(R) among aloe-emodin, rhein, chrysophanol, physcion and emodin were determined under different chromatographic conditions.</p><p><b>RESULT</b>Their RCFs showed no significant difference.</p><p><b>CONCLUSION</b>The RCFs among anthraquinones established by QAMS can be used as a constant in content determination of traditional Chinese medicines/patent traditional Chinese medicines.</p>
Subject(s)
Chromatography, High Pressure Liquid , Methods , Rheum , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To analyze the clinical data and result of voiding cystourethrography (VCUG) in high-risk children with vesicoureteral reflux (VUR) for better awareness of VUR, and to assess the usefulness of non-radioactive voiding ultrasonography (VUS) in the diagnosis of VUR.</p><p><b>METHOD</b>Ninety-three high-risk children with VUR who were hospitalized from July 2007 to April 2010 were studied. The study included 58 cases of urinary tract infection (UTI) and 35 cases of fetal or postnatal hydronephrosis detected on a B ultrasound scan. The results of urinalysis, urine culture, renal function, B ultrasound and VCUG were evaluated. Part of patients underwent VUS followed by VCUG immediately.</p><p><b>RESULT</b>(1) Sixty-two boys and 31 girls (aged 1 month to 11.5 years, mean age 2 years) were included. VUR was detected in 26 patients (28%) by VCUG. In terms of kidney-ureter units, VUR was detected in 36 of 186 kidney-ureter units, including 6 grade I, 3 grade II, 6 grade III, 15 grade IV and 6 grade V. (2) VUR was detected in 20 of 58 UTI patients (34.5%) by VCUG. The proportion of VUR in recurrent UTI group was 61.1%, much higher than that in first UTI group (22.5%). Thirteen of 20 VUR (65%) occurred in UTI patients under 1 year of age (M/F 10/3), with more bilateral VUR and severe grades of VUR than the older group. VUR was detected in 6 of 35 fetal or postnatal hydronephrosis patients (17.1%) by VCUG. (3) Twenty-two patients underwent both VUS and VCUG. VUR was detected in 4 patients and 6 kidney-ureter units by VCUG, while in 6 patients and 9 kidney-ureter units by VUS. Taking VCUG as the reference standard, VUS had a sensitivity of 100%, specificity of 92.1%, positive predictive value of 66.7%, and negative predictive value of 100%. There was a concordance rate of 93.2% between VUS and VCUG.</p><p><b>CONCLUSION</b>It is important to early screen VUR in UTI, fetal or postnatal hydronephrosis patients. There are more VUR, especially more bilateral VUR and severe grades of VUR, occurred in UTI patients under 1 year of age compared to older children. The incidence of VUR in recurrent UTI group was much higher than that in first UTI group. VUS is an accurate, reliable and radiation-free technique for the detection of VUR. It could be used to screen high-risk children for VUR and do the evaluation in the follow-up of VUR.</p>