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1.
Article in Chinese | WPRIM | ID: wpr-1030506

ABSTRACT

Objective The contents of 11 nucleosides and base components in 10 batches of samples from 5 provinces(cities)including Chongqing,Yunnan and Shaanxi were determined,and the differences in nucleosides and base components in Fritillaria taipaiensis were compared by chemometric analysis,and the quality was comprehensively evaluated,so as to provide a reference for the cultivation of excellent varieties and the selection of medicinal materials.Methods Nucleoside and base components were extracted from Fritillaria taipaiensis by ultrasonication in aqueous solutions,and the content of each component was determined by HPLC-DAD method.The origin was classified by principal component analysis(PCA)and hierarchical cluster analysis(HCA).Partial least squares discriminant analysis(PLS-DA)was used to determine the differentiated index components in Fritillaria taipaiensis.Then the differences in the contents of the index components among samples from different origins were compared.Results It was found that 11 nucleoside and base components differed significantly among different origins of Fritillaria taipaiensis.Principal component analysis and hierarchical cluster analysis indicated that all samples could be clustered into 4 categories.Five characteristic components,including uracil,cytosine,uridine,inosine,and adenosine,were identified by PLS-DA.The nucleosides and bases in samples from Chongqing and Hubei were relatively high,and the quality of the samples was comparatively superior.Conclusion This method is simple,reproducible,accurate and reliable.It has screened out the index nucleoside and base components in the identification of Fritillaria taipaiensis of different origins,which can be used to initially elucidate the differences of samples between different origins.Additionally,it can better reflect the quality of Fritillaria taipaiensis,and can provide reference for the selection of procurement origin and the quality control for Fritillaria taipaiensis.

2.
Journal of Clinical Hepatology ; (12): 880-883, 2024.
Article in Chinese | WPRIM | ID: wpr-1030779

ABSTRACT

Highly effective oral antiviral therapy with low drug resistance can strongly inhibit HBV replication; however, some patients may still have low-level viremia (LLV) after receiving entecavir, tenofovir disoproxil fumarate, tenofovir alafenamide, or tenofovir amibufenamide for 48 weeks or more. Multiple studies in China and globally show that LLV after antiviral therapy is closely associated with the progression of chronic hepatitis B liver fibrosis, the risk of decompensated liver cirrhosis and hepatocellular carcinoma, and the reduction in long-term survival rate. Therefore, this article reviews the development, risk factors, and clinical harm of LLV after first-line treatment with nucleos(t)ide analogues, as well as different treatment regimens, in order to provide a reference for the treatment of LLV in chronic hepatitis B patients in the future.

3.
Article in Chinese | WPRIM | ID: wpr-1006427

ABSTRACT

‍ ObjectiveTo investigate the serum level of HBV RNA in untreated or treatment-experienced patients with chronic hepatitis B (CHB) and the correlation between serum HBV RNA level and the duration of antiviral therapy with nucleos(t)ide analogues (NAs). MethodsA total of 300 patients with CHB who attended Department of Infectious Diseases in The First Affiliated Hospital of Shihezi University School of Medicine from February to July, 2022, were enrolled as subjects. Related clinical data were collected, and according to the duration of antiviral therapy, they were divided into untreated group with 73 patients, treatment duration ≤1 year group with 91 patients, and treatment duration >1 year group with 136 patients. Serum HBV RNA load, HBV DNA load, and HBsAg concentration were measured for all patients. The Mann-Whitney U test was used for comparison of continuous data between two groups, and the Kruskal-Wallis H test was used for comparison between multiple groups, further pairwise comparison using Bonferroni method; the chi-square test was used for comparison of categorical data; a Spearman correlation analysis was used to investigate the degree of correlation between various indicators. ResultsThe positive rate of HBeAg was 18.3%, and among the patients with negative HBV DNA, the patients with positive HBV RNA accounted for 44.1% (86/195). There was a significant difference in the distribution of the serum levels of HBV RNA, HBV DNA, and HBsAg between the positive HBeAg group and the negative HBeAg group (Z=10.740, 6.300, and 7.280, all P<0.05). There was a significant difference in the distribution of DNA level between the untreated group and the treatment duration ≤1 year group (P<0.05); there was a significant difference in the distribution of HBV RNA and HBV DNA levels between the untreated group and the treatment duration >1 year group (P<0.05); there was a significant difference in the distribution of HBV RNA, HBV DNA, and HBsAg levels between the treatment duration ≤1 year group and the treatment duration >1 year group (P<0.05). The correlation analysis between the duration of antiviral therapy and the levels of HBV RNA, HBV DNA, and HBsAg showed that the duration of antiviral therapy had an extremely weak negative correlation with the levels of HBV RNA and HBsAg (r=-0.247 and -0.138, both P<0.05) and a strong negative correlation with the level of HBV DNA (r=-0.771, P<0.001). There was a low degree of correlation between the serum level of HBV RNA and the serum levels of HBV DNA and HBsAg (r=0.360 and 0.442, both P<0.001). Further stratified analysis showed that in the untreated group, there was a strong positive correlation between HBV RNA and HBV DNA (r=0.752, P<0.001) and a moderate positive correlation between HBV RNA and HBsAg (r=0.559, P<0.001); in the treatment duration ≤1 year group, there was a low degree of positive correlation between HBV RNA and HBV DNA/HBsAg (r=0.396 and r=0.388, both P<0.001); in the treatment duration >1 year group, there was a low degree of positive correlation between HBV RNA and HBsAg (r=0.352, P<0.001). ConclusionSerum HBV RNA is negatively correlated with the duration of treatment with NAs, and the correlation of HBV RNA with HBV DNA and HBsAg gradually decreases with the increase in the duration of treatment. Therefore, it can be used as a supplementary indicator for monitoring the level of virologic response in CHB patients to a certain extent, with a relatively high accuracy in reflecting the level of viral replication in untreated patients.

4.
Chinese Herbal Medicines ; (4): 162-167, 2024.
Article in English | WPRIM | ID: wpr-1010742

ABSTRACT

OBJECTIVE@#Cultivated Cordyceps sinensis powder has been used as clinical drug and healthy food to nourish the lung and kidney, which solves the problem of serious shortage of wild C. sinensis. This study aims to explore the chemical components and compared their anti-fibrotic effects in cultivated C. sinensis.@*METHODS@#Nucleosides, sterols and polysaccharides were separated and purified from cultivated C. sinensis, and analyzed by high performance liquid chromatography, gas chromatography-mass spectrometry and chemical chromogenic methods, respectively. In high glucose-induced rat mesangial cell models, fibronectin and type 1 collagen were used as evaluation indicators.@*RESULTS@#There were 10 kinds of nucleosides and one sterol in cultivated C. sinensis. The contents of nucleosides, sterols and polysaccharides in the cultivated C. sinensis were close to 2%, 0.55% and 4.4%, respectively. Furthermore, nucleoside, sterol and polysaccharide components exhibited varying degrees of anti-fibrotic activity. The nucleoside components and sterol components inhibited the expression of extracellular matrix more effectively in the three main components.@*CONCLUSION@#Cultivated C. sinensis remains the similar compounds with the wild C. sinensis, and nucleosides and sterols may be the main active substances that contribute to its anti-fibrotic effects. The project of this study may provide valuable information on further optimization of more effective remedies with few side effects based on cultivated C. sinensis.

5.
Journal of Clinical Hepatology ; (12): 1070-1075, 2023.
Article in Chinese | WPRIM | ID: wpr-973194

ABSTRACT

Objective To investigate the ability of combined baseline serum markers, i.e., HBV DNA, HBV RNA, HBsAg, and HBcrAg, to predict HBeAg seroconversion in patients with HBeAg-positive chronic hepatitis B (CHB) treated by nucleos(t)ide analogues. Methods A retrospective analysis was performed for 83 HBeAg-positive patients selected as subjects from the prospective CHB follow-up cohort established by Difficult & Complicated Liver Diseases and Artificial Liver Center, Beijing YouAn Hospital, Capital Medical University, from June 2007 to July 2008, and the baseline serum levels of HBV DNA, HBV RNA, HBsAg, and HBcrAg were analyzed. The t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. The Spearman method was used for correlation analysis. A Cox regression model was established to calculate HBeAg seroconversion prediction score, and the time-dependent receiver operating characteristic curve was used to evaluate the ability of combined markers in predicting HBeAg seroconversion. The Kaplan-Meier method was used to calculate cumulative seroconversion rate in each group, and the Log-rank test was used for comparison between groups. Results For the 83 HBeAg-positive patients, the median follow-up time was 108 months, and 44.58%(37/83) of these patients achieved HBeAg seroconversion. Compared with the non-seroconversion group, the HBeAg seroconversion group had significantly lower baseline serum levels of HBV DNA [6.23(1.99-9.28) log 10 IU/mL vs 7.69(2.05-8.96) log 10 IU/mL, Z =-2.345, P =0.019] and HBV RNA [4.81(1.40-7.53) log 10 copies/mL vs 6.22(2.00-8.49) log 10 copies/mL, Z =-1.702, P =0.010], and there were no significant differences in the levels of HBsAg and HBcrAg between the two groups ( P > 0.05). The Cox regression equation constructed based on the above serum markers showed a median score of 0.95(range 0.37-3.45) for predicting HBeAg seroconversion. In the total population, the combined score was negatively correlated with HBsAg, HBV DNA, HBV RNA, and HBcrAg ( r =-0.697, -0.787, -0.990, and -0.819, all P < 0.001). Based on the median prediction score, the patients were divided into high HBeAg seroconversion group and low HBeAg seroconversion group; as for the prediction of HBeAg seroconversion rate at 36, 60, and 84 months, the high HBeAg seroconversion group had a seroconversion rate of 43.90%, 51.20%, and 63.10%, respectively, while the low HBeAg seroconversion group had a seroconversion rate of 9.60%, 17.00%, and 19.8%, respectively, and there was a significant difference between the two groups ( χ 2 =11.6, P < 0.001). Conclusion The combined prediction score based on baseline serum HBV markers can predict HBeAg seroconversion in CHB patients treated by nucleos(t)ide analogues.

6.
Zhongguo Zhong Yao Za Zhi ; (24): 114-125, 2023.
Article in Chinese | WPRIM | ID: wpr-970507

ABSTRACT

Galli Gigerii Endothelium Corneum(GGEC), the dried gizzard membrane of Gallus gallus domesticus is a Chinese medicinal material commonly used for digestion. However, due to the particularity of texture and composition, its active ingre-dients have not been clarified so far, and there is also a lack of quality evaluation indicators. In this study, UPLC-Q-TOF-MS was used to analyze the chemical components from the water extract of GGEC, and ten nucleosides were identified for the first time. HPLC fingerprints of the water extracts of GGEC were established and the content of seven nucleosides was determined. The fingerprint similarities of 40 batches of GGEC samples ranged from 0.765 to 0.959, indicating that there were great differences among the GGEC products processed with different methods. In addition, SPSS 22.0 and SIMCA 14.1 were used for hierarchical cluster analysis(HCA) and principal component analysis(PCA) on the 19 common peaks of the HPLC fingerprints of GGEC, and the 40 batches of samples were divided into three categories: raw GGEC, fried GGEC and vinegar-processed GGEC. Eight differential components in GGEC were marked by orthogonal partial least squares discrimination analysis(OPLS-DA), two of which were adenine and thymine. The results of content determination showed that the total content of the seven nucleosides in raw GGEC, fried GGEC and vinegar-processed GGEC were 182.5-416.8, 205.3-368.7, and 194.2-283.0 μg·g~(-1), respectively. There were significant differences in the content of hypoxanthine, thymine and thymidine among the GGEC products processed with different methods(P<0.05), which were graded in the order of fried GGEC>vinegar-processed GGEC>raw GGEC. This suggested that the content of hypoxanthine, thymine and thymidine tended to increase during the frying process, and the variation range might be related to the degree of heat exposure. The established methods in this study were simple and reproducible, and could be used for qualitative and quantitative analysis of GGEC and its processed pro-ducts. This study also provided reference for the establishment of quality standards of GGEC with chemical components as control index.


Subject(s)
Nucleosides , Drugs, Chinese Herbal/chemistry , Chromatography, High Pressure Liquid , Acetic Acid , Thymine , Thymidine , Water , Hypoxanthines
7.
Article in Chinese | WPRIM | ID: wpr-913156

ABSTRACT

Objective To investigate the difference in the prognosis of hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) caused by hepatitis recurrence after withdrawal of nucleos(t)ide analogues (NUC) and possible causes in HBeAg-positive versus HBeAg-negative chronic hepatitis B (CHB) patients. Methods A total of 108 CHB patients with HBV-ACLF caused by withdrawal of NUC who were admitted to The First Affiliated Hospital of Nanchang University from January 2017 to December 2018 were enrolled, and according to HBeAg status, these patients were divided into HBeAg-positive group with 57 patients and HBeAg-negative group with 51 patients. The two groups were compared in terms of sex, age, clinical manifestation, signs, levels of total bilirubin, direct bilirubin, alanine aminotransferase, aspartate aminotransferase, prothrombin time, activated partial thromboplastin time, prothrombin time/international normalized ratio, and HBV DNA quantification on admission, complications (including hepatic encephalopathy, hepatorenal syndrome, and spontaneous bacterial peritonitis), and prognosis of HBV-ACLF. In addition, 48 CHB patients with continuous NUC antiviral therapy for > 2 years and HBV DNA < 20 IU/mL were enrolled, and the serum level of HBV pgRNA was measured to investigate the possible causes of the difference in the prognosis of HBV-ACLF between the patients with different HBeAg statuses. The two-independent-samples t test was used for comparison of normally distributed continuous data between two groups, and the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data. Results For the 108 patients with HBV-ACLF caused by drug withdrawal and recurrence, the HBeAg-positive group had an improvement rate of 49.1% and the HBeAg-negative group had an improvement rate of 74.5%. The HBeAg-negative group had a significantly higher improvement rate than the HBeAg-positive group ( χ 2 =2.811, P =0.006). The HBeAg-positive group had a significantly higher level of HBV DNA than the HBeAg-negative group on admission ( t =-3.138, P =0.002). For the 48 CHB patients who achieved virologic response after long-term antiviral therapy, the HBeAg-positive group had a significantly higher HBV pgRNA load than the HBeAg-negative group ( H =2.814, P =0.049). Conclusion Compared with the HBeAg-positive CHB patients, HBeAg-negative CHB patients have a significantly better improvement rate of HBV-ACLF caused by hepatitis recurrence after withdrawal of NUC antiviral therapy. The difference in baseline HBV pgRNA level may be associated with the difference in the prognosis of HBV-ACLF in patients with different HBeAg statuses.

8.
Journal of Clinical Hepatology ; (12): 532-536, 2022.
Article in Chinese | WPRIM | ID: wpr-922947

ABSTRACT

Objective To investigate the efficacy of entecavir (ETV) versus tenofovir disoproxil fumarate (TDF) and the treatment measures for poor response in previously untreated chronic hepatitis B (CHB) patients with high viral load. Methods A total of 165 CHB patients who received antiviral therapy and met the inclusion criteria in Department of Infectious Diseases, The First Affiliated Hospital of Guangxi Medical University, from June 2016 to July 2021 were enrolled. The patients enrolled had a baseline HBV DNA level of > 6lg copies/ml and were previously untreated CHB patients who had used ETV or TDF for 48 weeks, and quantitative real-time PCR was used to measure HBV DNA. Virologic response rate was calculated after 48 weeks of treatment; a logistic regression analysis was used to investigate the influencing factors for the response of HBV DNA 40 U/L) at baseline, 89.2% (107/120) achieved an HBV DNA load of 30 years (77.8% vs 47.2%, 85.2% vs 66.7%). For the patients who did not achieve complete virologic response (HBV DNA ≥100 copies/mL) after 48 weeks of treatment, 87.9% (29/33) achieved complete virologic response after the original treatment regimen was prolonged for 48 weeks, and 100% (9/9) of the patients achieved complete virologic response after switching to or adding the first-line nucleos(t)ide analogues (NUCs) without cross-resistance sites with the original regimen for another 48 weeks. Conclusion The patients aged > 30 years should receive antiviral therapy as early as possible, regardless of viral load and ALT level, especially those with a family history of liver cirrhosis or hepatocellular carcinoma; the patients aged ≤30 years who have a normal ALT level and a high viral load should consider initiating antiviral therapy after providing informed consent. For the patients with poor response after 48 weeks of treatment, first-line NUCs without cross-resistance sites with the original regimen should be switched to or added in time.

9.
Journal of Clinical Hepatology ; (12): 1705-1709, 2022.
Article in Chinese | WPRIM | ID: wpr-941526

ABSTRACT

Chronic hepatitis B virus (HBV) infection is the main cause of viral hepatitis, liver cirrhosis, and primary liver cancer. At present, nucleos(t)ide analogues (NUC) and pegylated interferon α used in clinical practice cannot directly target covalently closed circular DNA, and it is difficult to achieve clinical cure of chronic hepatitis B patients; therefore, it is urgently needed to develop direct-acting antiviral agents targeting all stages of the HBV replication cycle. Capsid assembly modulator (CpAM) targets the assembly of viral capsids through various mechanisms, thereby exerting a direct-acting antiviral effect. Its combination with NUC should have a good synergistic antiviral effect, but the results of existing clinical trials have shown that chronic hepatitis B patients who received a limited course of antiviral therapy with CpAM and NUC all experienced off-therapy viral rebound. Based on the mechanism of action of these two types of drugs, this article provides a reasonable explanation for the above clinical trial results and points out that a longer course of antiviral therapy with CpAM and NUC may be needed in the future clinical trials with safe drug withdrawal as the end point of observation, so as to deplete or silence the pool of covalently closed circular DNA and increase the possibility of safe drug withdrawal in CHB patients. In addition, further studies are needed to explore antiviral therapeutic strategies with a combination of multiple targets.

10.
Journal of Clinical Hepatology ; (12): 1268-1274, 2021.
Article in Chinese | WPRIM | ID: wpr-877314

ABSTRACT

Nucleos(t)ide analogues (NAs), which are widely used as the first-line anti-hepatitis B virus (HBV) drugs in clinical practice, can effectively inhibit the replication of HBV DNA, significantly slow down disease progression in chronic hepatitis B (CHB) patients, and reduce the development of end-stage liver diseases such as liver failure and liver cancer. However, for some CHB patients receiving first-line NAs for 48 weeks or longer, serum HBV DNA is still persistently or intermittently higher than the lower detection of limit of sensitive nucleic acid detection reagents. After discussion by the authors, low-level viremia (LLV) is defined as follows: persistent LLV refers to the condition in which CHB patients, who receive entecavir, tenofovir disoproxil fumarate, or tenofovir alafenamide fumarate for ≥48 weeks, test positive for HBV DNA by two consecutive detections with sensitive quantitative PCR, with an interval of 3-6 months, but have an HBV DNA level of <2000 IU/ml; intermittent LLV refers to the condition in which patients test positive for HBV DNA intermittently by at least three consecutive detections with sensitive quantitative PCR, with an interval of 3-6 months, but have an HBV DNA level of <2000 IU/ml. For the diagnosis of LLV, the issues of poor compliance and drug-resistant mutations should be excluded. LLV might be associated with the increased risk of progression to liver fibrosis or hepatocellular carcinoma in patients with liver cirrhosis under NA treatment, but there are still controversies over whether the original treatment regimen with NAs should be changed after the onset of LLV. This article summarizes the incidence rate of LLV under NA treatment and the influence of LLV on prognosis and analyzes the possible mechanisms of the osnet of LLV, so as to provide a reference for the management of LLV in patients treated with NAs.

11.
Journal of Clinical Hepatology ; (12): 1793-1797., 2021.
Article in Chinese | WPRIM | ID: wpr-886332

ABSTRACT

ObjectiveTo investigate the population with an advantage of clinical cure previously treated with nucleos(t)ide analogues (NAs), and to provide more methods for clinicians in pursuing the clinical cure of hepatitis B. MethodsA total of 42 chronic hepatitis B patients with low-level HBsAg who received NAs treatment in Hebi Third People’s Hospital from October 2017 to October 2019 were enrolled as subjects and divided into combination treatment group (group A) and NA monotherapy group (group B). The 22 subjects in group A were treated with NAs combined with PEG-IFN antiviral therapy for 48 weeks, and some patients withdrew from PEG-IFN after 24 weeks and continued to receive NA monotherapy, while the 20 subjects in group B received NA antiviral therapy alone. Both groups were observed till week 48, and the five makers for hepatitis B were measured to evaluate clinical outcome. The t-test was used for comparison of continuous data between two groups, and the Fisher’s exact test was used for comparison of categorical data between two groups; a multivariate logistic regression analysis was used to perform a multivariate analysis. ResultsCompared with group B at the 48-week treatment endpoint, group A had significantly higher HBsAg clearance rate (45.5% vs 0, P<0.01) and HBsAg seroconversion rate (31.8% vs 0, P<0.01). The population with HBsAg <1000 IU/ml, <500 IU/ml, <100 IU/ml, and <10 IU/ml had an HBsAg clearance rate of 52.6%, 61.5%, 66.7%, and 100%, respectively, and the population with an HBsAg level of 500-1000 IU/ml, 100-500 IU/ml, 10-100 IU/ml, and <10 IU/ml had an HBsAg clearance rate of 33.3%, 50%, 40%, and 100%, respectively. The 4 patients with baseline HBsAg <10 IU/ml (accounting for 18.2% in group A) achieved clinical cure at week 12 of combined treatment, and after observation to week 48, 2 patients had an anti-HBs level of >100 IU/ml and 2 had an anti-HBs level of >1000 IU/ml. The multivariate logistic regression analysis of HBsAg clearance showed that age at the initiation of combined treatment affected HBsAg clearance (odds ratio [OR]=0.877, 95% confidence interval [CI]: 0.781-0.985, P=0.026), and most of the patients with HBsAg clearance had an age of 36-49 (44.20±4.49) years; baseline HBsAg level also had an impact on HBsAg clearance (OR=0.996, 95% CI: 0.992-1.000, P=0.050). ConclusionThe addition of interferon therapy in chronic hepatitis B patients with low-level HBsAg previously treated with NAs can significantly improve the clinical cure rate. The younger the age and the lower the HBsAg level, the shorter the duration of combined treatment. Age and baseline HBsAg level are more important than the duration and type of NA medication.

12.
China Pharmacy ; (12): 158-163, 2021.
Article in Chinese | WPRIM | ID: wpr-862637

ABSTRACT

OBJECTIVE:To establish fingerprint of Huafengdan yaomu ,and to determine the contents of 7 nucleosides in samples of different fermentation time. METHODS :HPLC method was adopted. The determination was performed on Pntulips BP-C18 column with mobile phase consisted of methanol-water (gradient elution )at the flow rate of 0.8 mL/min. The detection wavelength was set at 260 nm,and column temperature was 35 ℃. The sample size was 10 μL. Using xanthine as reference, HPLC fingerprint of 12 batches of Huafengdan yaomu was drawn. The similarity of samples were evaluated with Similarity Evaluation System of TCM Chromatographic Fingerprints (2012 edition). Common peaks were confirmed. The contents of uracil , hypoxanthine,xanthine,uridine,inosine,guanosine and thymidine were determined in samples of different fermentation time (0, 1,2,3,4 weeks)by the same method. RESULTS :There were 8 common peaks in 12 batches of Huafengdan yaomu ,with similarities ranging from 0.712 to 0.954;7 components were identified ,namely uracil ,hypoxanthine,xanthine,uridine,inosine, guanosine and thymidine. The linear ranges of mass concentrations of above 7 components in samples at different fermentation time were 0.87-8.7 μ g/mL (r=0.999 6), 4030 1.51-15.1 μg/mL(r=0.999 7),6.08-60.8 μg/mL(r=0.999 5), 号) 1.52-15.2 μg/mL(r=0.999 6),1.82-18.2 μg/mL(r=0.999 6), 1.48-14.8 μg/mL(r=0.999 6),1.63-16.3 μg/mL(r=0.999 3). The limits of quantification were 0.027 4,0.076 3,0.250 4,0.172 3,0.101 1,0.078 3,and 0.084 2 μ g/mL,and the detection limits were 0.008 7,0.025 5,0.007 9,0.084 1,0.035 7,0.026 9,0.027 5 μg/mL,respectively. RSDs of precision , repeatability and stability tests (12 h)were all less than 3%. The sample recovery rates were 94.16%-100.16%(RSD=2.24%,n= 6),93.87%-100.65%(RSD=2.67%,n=6),93.52%-99.66%(RSD=2.30%,n=6),93.67%-98.24%(RSD=1.89%,n=6), 96.00%-102.18%(RSD=1.96%,n=6),94.62%-101.54%(RSD=2.82%,n=6),97.72%-104.56%(RSD=2.97%,n=6). After fermentation for 0-4 weeks,the contents of the above 7 components and total nucleosides were 0.042-0.232,0.027-0.181, 0.039-0.651,0.026-0.225,0.034-0.111,0.009-0.124,0.079-0.099,0.647-1.292 mg/g,respectively. After fermentation for 1-4 weeks,the contents of uracil ,hypoxanthine,xanthine and total nucleosides were significantly increased ,compared with 0 week of fermentation;the contents of uridine ,inosine and guanosine were significantly lower than those in 0 weeks. CONCLUSIONS :The established fingerprint has strong characteristics and simple to operate ,which can be used for the quality control of Huafengdan yaomu;the content determination method is accurate and reliable ,and can be used to simultaneously determine the contents of 7 active nucleosides ;the content of nucleosides in Huafengdan muyao is affected by fermentation time.

13.
Article in Chinese | WPRIM | ID: wpr-905874

ABSTRACT

Objective:To compare the effects of different drying methods on the chemical constituents of Trichosanthis Fructus. Method:Trichosanthis Fructus was dried by means of air drying, sun drying, hot air drying (40, 60, 80 ℃) and variable temperature drying (50-80, 80-50 ℃). The contents of nucleosides and flavonoids in Trichosanthis Fructus peels and seeds treated by different methods were compared by high performance liquid chromatography (HPLC), mobile phase was acetonitrile-0.2% acetic acid aqueous solution (3∶7) (A)-acetonitrile (B) for gradient elution (0-15 min, 97-95%B; 15-30 min, 95%-90%B; 30-35 min, 90%-87%B; 35-40 min, 87%-86.5%B; 40-48 min, 86.5%-97%B; 48-50 min, 97%B), the detection wavelength was 260 nm, and the flow rate was 0.4 mL·min<sup>-1</sup>. Gas chromatography-ion mobility spectrometry (GC-IMS) was used to compare the changes of volatile components in the samples treated by different treatments. The volatile components were incubated on a SE-54 capillary column (0.32 mm×30 m, 0.25 μm) at 80 ℃ and 500 r·min<sup>-1</sup> for 15 min, the injection temperature was 85 ℃, the injection volume was 400 μL, the analysis time was 35 min, carrier gas was high purity nitrogen, the flow rate of carrier gas was 2.0 mL·min<sup>-1</sup>, the flow rate of drift gas was 150 mL·min<sup>-1</sup>, and the temperature of IMS detector was 45 ℃. Result:The contents of uridine, adenosine and adenine were higher after hot air drying at >50 ℃. Low temperature drying was conducive to maintaining the stability of cytidine, cytosine, rutin, luteolin and 2ʹ-deoxyadenosine. GC-IMS technology could realize the analysis and identification of Trichosanthis Fructus samples after different treatments. There were more volatile components after hot air drying at 80 ℃ and variable temperature drying. Conclusion:Hot air drying at 40 ℃ and 60 ℃ can retain nucleosides and flavonoids, and the volatile components are similar to those in traditional drying methods, which has the advantages of high efficient, controllable and suitable for industrial production.

14.
Zhongguo Zhong Yao Za Zhi ; (24): 3337-3348, 2021.
Article in Chinese | WPRIM | ID: wpr-887983

ABSTRACT

A high performance liquid chromatography( HPLC) method was established for the fast,and precise determination of ten nucleosides in Fritillariae Cirrhosae Bulbus and its counterfeits. Then multivariate statistical analyses,such as clustering analysis,principal component analysis( PCA),and Fisher' s linear discriminant analysis( LDA),were conducted to establish a discriminant function model for an integrated analysis. The results indicated that data acquisition time of a single sample was shortened within 16 min by the HPLC method. In the range of 5-1 000 mg·kg~(-1),the mass concentrations of all nucleosides exhibited good linear relationships with the corresponding peak areas( R2> 0. 999). The spiked recoveries were in the range of 93. 83%-108. 9% with RSDs of0. 12%-1. 3%( n = 5). The limit of quantitation( LOQ) was 0. 98-4. 13 mg·kg~(-1). As revealed by the clustering analysis,Fritillariae Cirrhosae Bulbus and the counterfeits could be discriminated into two clusters based on the content of nucleosides. Fisher's LDA could achieve this discrimination,while PCA dimension reduction failed. The accuracy of the discriminant function model established on the screened characteristic indicators reached 97. 5%. The present study proposed a new identification method of Fritillariae Cirrhosae Bulbus with one-dimensional indicators,which is simple,accurate,and reliable. It can provide a scientific basis for further optimizing the identification techniques for Fritillariae Cirrhosae Bulbus and inspiration for quality control strategy development of Chinese medicinal materials.


Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Fritillaria , Nucleosides , Plant Roots
15.
Zhongguo Zhong Yao Za Zhi ; (24): 3494-3503, 2021.
Article in Chinese | WPRIM | ID: wpr-888000

ABSTRACT

In order to reveal the regional characteristics of nucleosides and amino acids in Elaphuri Davidiani Cornu,39 samples of Elaphuri Davidiani Cornu collected from 4 different regions were analyzed by UPLC-QTRAP ~®/MS2 method followed by orthogonal partial least-squares discrimination analysis( OPLS-DA) and cluster analysis( CA). The results showed all the samples contained abundant nucleosides and amino acids,with the total content of 45. 09 μg·g~(-1) and 634. 80 μg·g-1,respectively. The samples presented significant regional differences in the contents of individual components,and the main differential components included Ura,Hpro,Thr,Glu,G5 P,2'-dG,Adeno,Met,Ade,Gln,Orni,Phe,2'-dA,Hit,Lys,and Ile. Among them,Ura,Met,Glu,and Ile had the highest content in the samples from Dafeng in Jiangsu,Qinhu in Jiangsu,Beijing,and Shishou in Hubei,respectively. OPLS-DA and CA demonstrated that all the samples of Elaphuri Davidiani Cornu could be divided into three categories,reflecting the regional characteristics. The results indicated that the accumulation of nucleosides and amino acids in Elaphuri Davidiani Cornu was closely related to its habitat,providing a useful reference for the research on the quality formation,quality evaluation and control,as well as the comprehensive utilization of Elaphuri Davidiani Cornu. The findings suggested that the content factors of Ura,Met,Glu,and Ile could be included into the quality standard system of Elaphuri Davidiani Cornu as the characteristics of medicinal materials from different regions.


Subject(s)
Amino Acids , Beijing , Cornus , Nucleosides
16.
Zhongguo Zhong Yao Za Zhi ; (24): 2912-2922, 2021.
Article in Chinese | WPRIM | ID: wpr-888029

ABSTRACT

The dried fruit body of Phylloporia ribis(Hymenochaetaceae), which prefers to live on the stumps of Lonicera japonica(Caprifoliaceae), has a variety of activities, whereas its pharmacodynamic material basis is not completely clear and there are few reports on its quality control and evaluation. In this study, an UPLC-Q-TOF-MS method was used to analyze the nucleosides and nucleobases in P. ribis and a HPLC method was established for simultaneous determination of 10 nucleosides and nucleobases. MS and MS/MS data were acquired in positive ion mode. Based on the data comparison of the sample and the reference substance, the literature data and the compound databases of ChemSpider and PubChem, 18 nucleosides and nucleobases were identified qualitatively from the water extract of P. ribis for the first time. After optimization, the HPLC was performed using a Welch Ultimate AQ C_(18) column(4.6 mm×250 mm, 5 μm) by gradient elution with acetonitrile and water as mobile phase, the flow rate of 1.0 mL·min~(-1), the detection wavelength of 260 nm, and the column temperature of 30 ℃. Through the investigation of the extraction method, solvent and time, it was determined that the test solution should be obtained by cold water extraction for 18 h. At the present HPLC conditions, 10 components of uracil, cytidine, hypoxanthine, uridine, thymine, inosine, guanosine, 2'-deoxyinosine, 2'-deoxyguanosine and thymidine could be well separated(R > 1.5) and showed good linearity(r > 0.999 9) in the concentration ranges of 0.247-24.7, 0.283-28.3, 0.273-27.3, 0.256-25.6, 0.257-25.7, 0.318-31.8, 0.245-24.5, 0.267-26.7, 0.250-25.0 and 0.267-26.7 mg·L~(-1), respectively. The average reco-veries of 10 components were 95.78%-104.5%, and the RSDs were 2.2%-5.2%(n=6). The contents of 10 nucleosides and nucleobases in different samples of P. ribis varied greatly, which were 0.021-0.122, 0.004-0.029, 0.014-0.226, 0.009-0.442, 0.003-0.014, 0.002-0.146, 0.007-0.098, 0-0.054, 0.005-0.069, 0.004-0.081 and 0.072-1.28 mg·g~(-1) for uracil, cytidine, hypoxanthine, uridine, thymine, inosine, guanosine, 2'-deoxyinosine, 2'-deoxyguanosine, thymidine and total 10 components, respectively. These results demonstrated that the components had significant differences in the internal quality, and good quality control was needed to ensure the medical efficacy. This study provides a scientific basis for the discovery of pharmacodynamic ingredients, quality control and evaluation of P. ribis.


Subject(s)
Basidiomycota , Chromatography, High Pressure Liquid , Guanosine , Nucleosides , Tandem Mass Spectrometry
17.
China Pharmacy ; (12): 1747-1751, 2021.
Article in Chinese | WPRIM | ID: wpr-882147

ABSTRACT

OBJECTIVE:To establish the fingerprint of Bombyx mori and the method for the content determination of multi- components,and to provide reference for comprehensive quality evaluation of B. mori . METHODS :Using 18 batches of B. mori from different producing areas as samples ,HPLC method was used. The column was Shiseido CAPCELL PAK C 18 AQ S 5 with mobile phase consisted of methanol- 0.05 mol/L potassium dihydrogen phosphate solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelength was 260 nm,and column temperature was set at 30 ℃. The sample size was 10 μL. HPLC fingerprint analysis and similarity evaluation were performed by using TCM Chromatogram Fingerprint Similarity Evaluation System(2012 edition),and the chromatographic peak was identified by comparing with the chromatogram of reference substance. The contents of 4 nucleosides as uracil ,guanine,xanthine,uridine were determined . RESULTS :A total of 16 common peaks were identified in HPLC fingerprint for 18 batches of B. mori ,and peaks 3,6 ,7 and 8 were identified as uracil ,guanine,xanthine and uridine. The similarity of sample chromatogram with control fingerprint were 0.912-1.000. The linear range of uracil ,guanine, xanthine and uridine were 5.34-534,5.28-528,5.06-506,5.195-519.5 μg/mL(r≥0.999 8). The limits of detection were 0.032 4, 0.032 0,0.030 7,0.031 2 μg/mL,and the limits of quantitation were 0.106 8,0.105 6,0.101 2,0.103 0 μg/mL. RSDs of precision,reproducibility and stability tests (24 h)were all lower than 1.00%(n=6). Average recoveries were 100.15%-102.95%, and RSD s were all lower than 2.00%(n=9). The content determination results showed that the content of uracil ,guanine, xanthine and uridine of B. mori from different producing areas were 0.41%-2.46%,0.37%-1.98%,0.72%-2.63%,0.94%-3.67%, respectively. CONCLUSIONS :Established HPLC fingerprint and content determination method of 4 nucleosides were specific , accurate and reliable ,which can be used for the quality evaluation and control of B. mori .

18.
Article in Chinese | WPRIM | ID: wpr-873166

ABSTRACT

Objective:To study the water soluble chemical constituents in rhizoma of Acorus tatarinowii and transformation pathway of nucleosides in the process of water extraction. Method:Compounds were isolated and purified by column chromatography on macroporous resin,Sephadex LH-20,ODS and preparative HPLC. Their structures were identified on the basis of physicochemical properties and spectral data. Nucleosides were identified from aqueous extract of A. tatarinowii,and their stability was investigated by HPLC. The possible transformation pathways of nucleosides in aqueous extract of A. tatarinowii were studied by nucleotide addition test. Result:Eleven compounds including four nucleosides,four phenylpropanoids,two alkaloids and a furfural were isolated,and identified as uridine(1),adenine(2),guanosine(3),adenosine(4),5-hydroxymethylfurfural(5),5-(hydroxymethyl)-1H-pyrrole-2-carboxaldehyde(6),(threo)1',2'-dihydroxyasarone(7),(erythro)1',2'-dihydroxyasarone(8),acoraminol A(9),acoraminol B(10),and tatarine A(11). The chromatographic peaks of compounds 1-4 and cytidine were identified from aqueous extract of A. tatarinowii by HPLC. After ultrasonic extraction for 0.5 h,the stability of nucleosides in water was poor. After ultrasonic extraction for 3 h or refluxing extraction for 0.5 h,the stability of nucleosides in water was good. Four transformation pathways including 5'-cytidylic acid→cytidine,uridine monophosphate→uridine,guanosine monophosphate,guanosine and adenosine-5'-monophosphate,adenosine 5'-diphosphate,adenosine 5'-triphosphorate,adenosine,adenine might exist in water extract of A. tatarinowii. Conclusion:Compounds 1-4 and 6 were isolated from the genus Acorus for the first time. These compounds further enriched the chemical constituents of A. tatarinowii. The stability and transformation pathway of nucleosides in A. tatarinowii provides reference data for the analysis of nucleosides in A. tatarinowii and other traditional Chinese medicine.

19.
Zhongcaoyao ; Zhongcaoyao;(24): 3147-3156, 2020.
Article in Chinese | WPRIM | ID: wpr-846351

ABSTRACT

Objective: To analyze and identify the chemical constituents from Lindley eupatorium by using UPLC-Q-TOF/MS. Methods: The separation was performed on Waters Acquity UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm) column with gradient elution of 0.1% formic acid (A)-acetonitrile (B), the flow rate was 0.2 mL/min. The column temperature was set at 35 ℃. The MS analysis was based on information associated mode (IDA), and positive and negative ions were collected respectively. Results: A total of 26 compounds in L. eupatorium were identified by PeakView, combined with the mass spectrometry data of each chromatographic peak in the database, and the cleavage law of secondary fragment of each peak of which11 compounds were first reported for L. eupatorium. The main chemical constituents included flavonoids, nucleosides, alkaloids, phenylpropanoid, sesquiterpenoids, coumarins, polyols, etc. Conclusion: The method is accurate, reliable and effficient, which is suitable for rapid identification of ingredients in L. eupatorium, which provides a reference for clarify its efficacy and material basis.

20.
Journal of Clinical Hepatology ; (12): 1358-1361, 2020.
Article in Chinese | WPRIM | ID: wpr-822190

ABSTRACT

Nucleos(t)ide analogues (NAs) are effective inhibitors for HBV replication and have become the preferred antiviral regimen for most patients with chronic hepatitis B (CHB). HBeAg seroconversion is an important index used to evaluate the durability and efficacy of antiviral therapy in HBeAg-positive CHB patients. The search for biomarkers that can predict HBeAg clearance or seroconversion after NAs treatment plays an important role in the selection of antiviral drugs, the adjustment of treatment regimens, and the achievement of individualized treatment. This article reviews the value of related markers, including HBV DNA, HBV RNA, anti-HBc, and HBcrAg, in predicting HBeAg clearance or seroconversion in CHB patients treated with NAs.

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