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As a local variety of medicinal material, Citri Trifoliatae Fructus is widely used in many places, whereas its harvest time remains unclear. Therefore, studying its harvest time can make more reasonable use of this medicinal material. In this study, we determined the flavonoids content and compared the color of Citri Trifoliatae Fructus harvested in different time, aiming to guide the harvest of this medicinal material. The fresh fruits of Citrus trifoliata were collected from Xinxiang city, Henan province, graded according to the diameter range, and then dried. The contents of isonaringin, naringen, and poncirin in Citri Trifoliatae Fructus were determined by HPLC, and the color values of the samples were detected by electronic eye. The correlation analysis of the obtained data was carried out to explore the relationships of color and diameter with quality. The results showed that the contents of isonaringin, naringen, and poncirin varied significantly in different harvest time, within the ranges of 0.21-1.20, 2.21-11.59, and 3.73-23.16 mg·g~(-1), respectively. With the delay of harvest time, Citri Trifoliatae Fructus showed the color changing from green to yellow, gradually increased diameter, and gradually decreased contents of isonaringin, naringen, and poncirin. The contents of isonaringin, naringen, and poncirin were negatively correlated with the degree of red and green(a~*) and positively correlated with the degree of yellow and blue(b~*). The contents of naringen and poncirin had significantly negative correlations with the diameter. This study indicates that the quality of Citri Trifoliatae Fructus can be judged by its diameter and skin color, which provides a theoretical basis for the rational harvest of this medicinal material.
Subject(s)
Chromatography, High Pressure Liquid , Citrus , Drugs, Chinese Herbal , Electronics , Fruit , TechnologyABSTRACT
Background@#Medicines for the treatment of 2019-novel coronavirus (2019-nCoV) infections are urgently needed. However, drug screening using live 2019-nCoV requires high-level biosafety facilities, which imposes an obstacle for those without such facilities or 2019-novel coronavirus (2019-nCoV). This study aims to repurpose the clinically approved drugs for the treatment of coronavirus disease 2019 (COVID-19) in a 2019-nCoV related coronavirus model.@*Methods@#A 2019-nCoV related pangolin coronavirus GX_P2V/pangolin/2017/ Guangxi was described. Whether GX_P2X uses angiotensin-converting enzyme 2 (ACE2) as the cell receptor was investigated by using small interfering RNA (siRNA) -mediated silencing of ACE2. The pangolin coronavirus model was used to identify drug candidates for treating 2019-nCoV infection. Two libraries of 2406 clinically approved drugs were screened for their ability to inhibit cytopathic effects on Vero E6 cells by GX_P2X infection. The antiviral activities and antiviral mechanisms of potential drugs were further investigated. Viral yields of RNAs and infectious particles were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and plaque assay, respectively.@*Results@#The spike protein of coronavirus GX_P2V shares 92.2% amino acid identity with that of 2019-nCoV isolate Wuhan-hu-1, and uses ACE2 as the receptor for infection just like 2019-nCoV. Three drugs-cepharanthine (CEP), selamectin and mefloquine hydrochloride exhibited complete inhibition of cytopathic effects in cell culture at 10 μmol/L. CEP demonstrated the most potent inhibition of GX_P2V infection, with a concentration for 50% of maximal effect [EC50] of 0.98 μmol/L. The viral RNA yield in cells treated with 10 μmol/L CEP was 15,393-fold lower than in cells without CEP treatment ([6.48±0.02]×10-4 vs. 1.00±0.12, t=150.38, P<0.001) at 72 h post-infection (p.i.). Plaque assays found no production of live viruses in media containing 10 μmol/L CEP at 48 h p.i. Furthermore, we found CEP has potent antiviral activities against both viral entry (1.00±0.37 vs. 0.46±0.12, t=2.42, P<0.05) and viral replication (1.00±0.43 vs. [6.18±0.95]×10-4, t=3.98, P<0.05).@*Conclusions@#Our pangolin coronavirus GX_P2V is a workable model for 2019-nCoV research. CEP, selamectin and mefloquine hydrochloride are potential drugs for treating 2019-nCoV infection. Our results strongly suggest that CEP is a wide-spectrum inhibitor of pan-betacoronavirus, and clinical trial of CEP for treatment of 2019-nCoV infection is warranted.
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BACKGROUND@#Medicines for the treatment of 2019-novel coronavirus (2019-nCoV) infections are urgently needed. However, drug screening using live 2019-nCoV requires high-level biosafety facilities, which imposes an obstacle for those institutions without such facilities or 2019-nCoV. This study aims to repurpose the clinically approved drugs for the treatment of coronavirus disease 2019 (COVID-19) in a 2019-nCoV-related coronavirus model.@*METHODS@#A 2019-nCoV-related pangolin coronavirus GX_P2V/pangolin/2017/Guangxi was described. Whether GX_P2V uses angiotensin-converting enzyme 2 (ACE2) as the cell receptor was investigated by using small interfering RNA (siRNA)-mediated silencing of ACE2. The pangolin coronavirus model was used to identify drug candidates for treating 2019-nCoV infection. Two libraries of 2406 clinically approved drugs were screened for their ability to inhibit cytopathic effects on Vero E6 cells by GX_P2V infection. The anti-viral activities and anti-viral mechanisms of potential drugs were further investigated. Viral yields of RNAs and infectious particles were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and plaque assay, respectively.@*RESULTS@#The spike protein of coronavirus GX_P2V shares 92.2% amino acid identity with that of 2019-nCoV isolate Wuhan-hu-1, and uses ACE2 as the receptor for infection just like 2019-nCoV. Three drugs, including cepharanthine (CEP), selamectin, and mefloquine hydrochloride, exhibited complete inhibition of cytopathic effects in cell culture at 10 μmol/L. CEP demonstrated the most potent inhibition of GX_P2V infection, with a concentration for 50% of maximal effect [EC50] of 0.98 μmol/L. The viral RNA yield in cells treated with 10 μmol/L CEP was 15,393-fold lower than in cells without CEP treatment ([6.48 ± 0.02] × 10vs. 1.00 ± 0.12, t = 150.38, P < 0.001) at 72 h post-infection (p.i.). Plaque assays found no production of live viruses in media containing 10 μmol/L CEP at 48 h p.i. Furthermore, we found CEP had potent anti-viral activities against both viral entry (0.46 ± 0.12, vs.1.00 ± 0.37, t = 2.42, P < 0.05) and viral replication ([6.18 ± 0.95] × 10vs. 1.00 ± 0.43, t = 3.98, P < 0.05).@*CONCLUSIONS@#Our pangolin coronavirus GX_P2V is a workable model for 2019-nCoV research. CEP, selamectin, and mefloquine hydrochloride are potential drugs for treating 2019-nCoV infection. Our results strongly suggest that CEP is a wide-spectrum inhibitor of pan-betacoronavirus, and further study of CEP for treatment of 2019-nCoV infection is warranted.
Subject(s)
Humans , Betacoronavirus , Genetics , Cell Line , Clinical Laboratory Techniques , Coronavirus Infections , Diagnosis , Drug Therapy , Drug Approval , Pandemics , Pneumonia, Viral , Diagnosis , Drug Therapy , RNA, Small Interfering , Genetics , Real-Time Polymerase Chain Reaction , Viral LoadABSTRACT
Objective To extract and isolate the lectin from Tibetan medicine Auricularia auricula. Methods The lectin from Tibet A. auricula was separated by sephadex column chromatography and ion exchange chromatography, and then its relative molecular weight was determinated by using matrix assisted laser desorption ionization time of flight mass spectrometry (5 800 MALDI-TOF/TOF). N-terminal sequential detection results suggested the amino acid sequences by comparing with the UniProt database. Results A kind of lectin was obtained from the separation of Tibet A. auricula with the molecular weight of of 18 913.22, in which the N- terminal amino acid sequence was detected as ITAPTTTSSAATE by the full automatic protein polypeptide sequencing instrument. The amino acid sequences of four peptide fragments were QIDAERK, TNHSVVTWNDK, RLNFTAGNPFPR, and VRELEQQVDSMTK. Conclusion These sequences are not found in the existing protein database of A. auricula, indicating that the isolated lectin should be a new type protein and it is confirmed as a new kind of lectin.
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BACKGROUND:The thickening of living bone is an important natural rule,and genetic,nutritional and endocrine factors play critical roles in the bone thickening;however,these factors are site-blind.OBJECTIVE:To analyze the mechanism underlying the thickening of the long bone.METHODS:Clinical cases and literature were analyzed.Medline and CNKI databases were retrieved using the keywords of "bone growth in width,skeletal thickening,bone thickening,periosteal apposition" in English and Chinese,respectively.Totally 12 eligible articles were included for result analysis.RESULTS AND CONCLUSION:The mechanical environment of the bone plays a key role in the bone remodeling,which is a site-specific process.The thickening of the long bone is completed through the periosteal apposition,and the coupling effect of stress-angiogenesis-osteogenesis induced by overloading is the pathological basis of bone thickening.The periosteal apposition and periosteal resorption belong to a lifelong dynamic process,and the bending stress is the most important factor to maintain the periosteal osteogenesis.If the bone is shielded from bending stress to a certain extent by the metal fixators,it may lead to bone resorption and thinning,nonunion and other serious consequences.Therefore,the strength of the fixators should match the patient body mass and the thickness of the bone.The marrow cavity is a natural low stress region,so intramedullary fixation theoretically has little effect on the width growth of bone.
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<p><b>BACKGROUND</b>Unregulated commercial blood/plasma collection among farmers occurred between 1992 and 1995 in central China and caused the second major epidemic of human immunodeficiency virus type 1 (HIV-1) infection in China. It is important to characterize HIV-1-infected former blood donors and to study characteristics associated with disease progression for future clinical intervention and vaccine development.</p><p><b>METHODS</b>A cross-sectional study was performed on HIV-1-infected former blood donors (FBDs) and age-matched HIV-seronegative local residents. Demographic, epidemiologic, clinical and key laboratory data were collected from all study participants. Both unadjusted and adjusted multivariate linear regressions were employed to analyze the association of the decrease of CD4(+) T-cell counts with other characteristics.</p><p><b>RESULTS</b>Two hundred and ninety-four HIV-1-infected FBDs and 59 age-matched HIV-seronegative local residents were enrolled in this study. The unregulated blood/plasma collection occurred more than a decade (10.8 - 12.8 years) ago, which caused the rapid spread of HIV-1 infection and the high prevalence of co-infection with hepatitis C virus (HCV, 89.5%); hepatitis B virus (HBV) co-infection was observed in only 11 HIV(+)participants (3.7%). Deterioration in both clinical manifestation and laboratory parameters and increase of viral loads were observed in parallel with the decrease of CD4(+) T-cell counts. The decrease of total lymphocyte counts (P < 0.001) and hemoglobin levels (P < 0.001) and the appearance of dermatosis (P = 0.03) were observed in parallel with the decrease of CD4(+) T-cell counts whereas viral loads (P < 0.001) and CD8(+) T-cell counts (P = 0.01) were inversely associated with CD4(+) T-cell counts.</p><p><b>CONCLUSIONS</b>Co-infection with HCV but not HBV is highly prevalent among HIV-1-infected FBDs. CD4(+) T-cell counts is a reliable indicator for disease progression among FBDs. Total lymphocyte counts, hemoglobin level and appearance of dermatosis were positively associated with CD8(+) T-cell counts and viral loads were inversely associated with the decreased CD4(+) T-cell counts.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Blood Donors , China , Epidemiology , Cross-Sectional Studies , HIV Infections , Epidemiology , Allergy and Immunology , HIV-1 , Hepatitis CABSTRACT
<p><b>OBJECTIVE</b>To study the impact of gender factor on the candidate gene study of essential hypertension (EH).</p><p><b>METHODS</b>The polymerase chain reaction (PCR) was performed to analyze the ACE gene I/D polymorphism of hypertensive patients (50 men and 50 women) and normal controls (50 men and 50 women). The investigation was further focused on possible influence of sex proportion on the conclusion of this kind of research.</p><p><b>RESULTS</b>The frequency of DD genotype in male hypertensive patients is significantly higher than that in male normal controls (chi(2) = 6.98, P = 0.004). The frequency of D allele in male EH group is significantly higher than that of male normal controls (chi(2) = 6.87, P = 0.009), while no significant difference was observed for II and ID genotype between male EH group and control group (P > 0.05). For female EH group and normal controls, there were no significant differences in frequency of genotype and allele (P > 0.05), the distribution ratio of DD genotype in male EH group is significantly different from that of female EH group (chi(2) = 4.06, P = 0.044). Furthermore, males with DD genotype in EH group had higher SBP and PP than that of males with II and ID genotype (P < 0.05). However, there was no significant difference in DBP in all three genotypes (P > 0.05). At the same time, there was no difference in SBP, DBP and PP (P > 0.05) between II and ID genotype in male EH group. In female hypertensive patients, there was no significant difference in SBP, DBP and PP between all three genotypes (P > 0.05).</p><p><b>CONCLUSIONS</b>The relationship between DD genotype in male and EH (especially SBP and PP) is closer than any other genotype-EH relationships in both male and female. The gender factor, as a probable confounding factor, can affect many candidate gene studies of essential hypertension including ACE gene I/D polymorphism, and thus biases the conclusion.</p>