OBJECTIVE To establish the HPLC fingerprint and content determination of five components in Ranunculus sceleratus L.. METHODS The separation was developed on an Agilent ZORBAX SB C18 chromatographic (4.6 mm×250 mm, 5 μm)column by gradient elution with methanol(A)-0.1 % phosphoric acid aqueous solution(B) as mobile phase to establish HPLC fingerprint of Ranunculus sceleratus L.. Combined with similarity evaluation, cluster analysis, principal component analysis, and orthogonal partial least squares-discriminant analysis, the quality of 13 batches of Ranunculus sceleratus L. was evaluated. RESULTS Thirteen batches of Ranunculus sceleratus L. samples were calibrated with 20 common peaks, of which 5 common peaks were identified, and the similarity ranged from 0.874 to 0.984. The results of cluster analysis and principal component analysis were basically the same, indicating that there might be differences in the content of chemical components of Ranunculus sceleratus L. in different regions. Protocatechuic aldehyde, caffeic acid, ferulic acid, hyperoside and isoquercitrin were determined in thirteen batches of Ranunculus sceleratus L., and their contents were 0.016−0.035, 0.010−0.070, 0.010−0.029, 0.016−0.051, 0.028−0.086 mg·g–1, respectively. CONCLUSION The established HPLC fingerprint and content determination method is simple, stable, and reliable, which can be used for qualitative analysis and provide reference to quality evaluation and resource utilization of Ranunculus sceleratus L..

Objective:To compare and evaluate the quality of wild and different cultivation methods of Sanghuang porus vaninii (Ljub.) L.W. Zhou & Y.C. Dai through analysis on UPLC characteristic atlas and multi-component content determination results. Methods:UPLC was used to establish the characteristic chromatogram and multi-component content determination method of Sanghuang porus vaninii (Ljub.) L.W. Zhou & Y.C. Dai, and clustering analysis, orthogonal partial least squares - discriminant analysis method were used for chemical pattern recognition analysis. Results:The results showed that there were 10 common peaks in 18 batches of Sanghuang porus vaninii (Ljub.) L.W. Zhou & Y.C. Dai. Five components were identified, erythrothioneine(peak 1), protocatechuic acid (peak 2), protocatechualdehyde (peak 3), caffeic acid (peak 4) and Hispidin (peak 5). HCA and OPLS-DA could distinguish Sanghuang porus vaninii (Ljub.) with different cultivation methods. Conclusion:Sanghuang porus vaninii (Ljub.) L.W. Zhou & Y.C. Dai in wood is closer to wild Sanghuang porus vaninii (Ljub.) L.W. Zhou & Y.C. Dai than in substitute cultivation. The UPLC characteristic atlas and multi-component content determination method established in this study can provide reference for the quality evaluation of Sanghuang porus vaninii (Ljub.) L.W. Zhou & Y.C. Dai.

BACKGROUND:Many clinical research observations have indicated a close association between rheumatoid arthritis and osteoporosis as well as bone mineral density(BMD).However,it remains unclear whether there is a causal genetic relationship between rheumatoid arthritis and the development of osteoporosis and alterations of BMD. OBJECTIVE:To assess the potential causal relationship between rheumatoid arthritis and osteoporosis as well as BMD using a two-sample Mendelian randomization approach,provide meaningful insights from a genetic perspective into the underlying mechanisms and offer a reference for early prevention of osteoporosis and improvement in the progression of the disease. METHODS:We conducted a study using data from publicly available genome-wide association studies databases to identify single nucleotide polymorphisms associated with rheumatoid arthritis as instrumental variables(P＜5×10-8).The main outcomes of the study included osteoporosis and BMD at five different sites,including total body BMD,lumbar spine BMD,femoral neck BMD,heel BMD,and forearm BMD.The inverse variance-weighted method was used as the primary analysis method to evaluate causal effects.Weighted median,simple median,weighted mode and MR-Egger regression were used as supplementary analyses.Causal relationships between rheumatoid arthritis and the risk of osteoporosis and BMD were assessed using odds ratios(OR)and 95%confidence intervals(CI).Heterogeneity was assessed using Cochran's Q test and horizontal pleiotropy was evaluated using MR-Egger intercept tests. RESULTS AND CONCLUSION:The inverse variance-weighted analysis demonstrated a positive association between genetically predicted rheumatoid arthritis and osteoporosis(OR=1.123,95%CI:1.077-1.171;P=4.02×10-8).Heterogeneity test(P=0.388)indicated no significant heterogeneity among the single nucleotide polymorphisms.MR-Egger intercept(P=0.571)tests did not detect horizontal pleiotropy,and sensitivity analysis showed no evidence of bias in the study results.There was no causal relationship between rheumatoid arthritis and BMD at the five different sites.The total body BMD(OR=1.000,95%CI:0.988-1.012;P=0.925),lumbar spine BMD(OR=0.999,95%CI:0.982-1.016;P=0.937),femoral neck BMD(OR=1.001,95%CI:0.986-1.016;P=0.866),heel BMD(OR=0.996,95%CI:0.989-1.004;P=0.419),and forearm BMD(OR=1.063,95%CI:0.970-1.031;P=0.996)indicated no significant association.MR-Egger intercept analysis did not detect potential horizontal pleiotropy(total body BMD:P=0.253;lumbar spine BMD:P=0.638;femoral neck BMD:P=0.553;heel BMD:P=0.444;forearm BMD:P=0.079).Rheumatoid arthritis may contribute to the development of osteoporosis through the interaction between chronic inflammation and bone formation,resorption,and absorption.Additionally,the use of glucocorticoids and the presence of autoantibodies(such as anti-citrullinated protein antibody)in patients with rheumatoid arthritis showed associations with osteoporosis.Future research should focus on monitoring systemic inflammatory markers,standardized use of glucocorticoids,and regular screening for osteoporosis risk in patients with rheumatoid arthritis.

Objective:To establish the fingerprint and stoichiometric analysis mode of Ultra Performance Liquid Chromatography Tandem Quadrupole Time-of-Flight Mass Spectrometry (UPLC-Q-TOF-MS) of Fritillaria anhuiensis Bulbus, so as to provide reference for its quality evaluation and standard formulation. Methods:By setting the CORTECS C18 column at 4.6 mm×150 mm, 2.7 μm with the mobile phases of acetonitrile-0.1% formic acid and 10 mmol/L aqueous ammonium formate for gradient elution at a flow rate of 0.4 ml/min and an injection volume of 2.0 μl. The TCM fingerprint similarity evaluation system (2012 version) was used to evaluate 9 batches of Fritillaria anhuiensis Bulbus samples. By using Liquid Chromatography-Mass Spectrometry (LC-MS) technique to make quantity analysis and by combining cluster analysis, principal component analysis and orthogonal least squares-discriminant analysis to make overall quality evaluation. Results:The fingerprint profiles of different batches of Fritillaria anhuiensis Bulbus were established and 21 common peaks were identified, and 12 of them were initially identified. Cluster analysis, principal component analysis and orthogonal least squares-discriminant analysis were used to cluster the nine batches of Fritillaria anhuiensis Bulbus into three categories. Conclusion:The fingerprint established in this study combined with the chemical pattern recognition method are highly sensitive and specific, which could reflect the overall characteristics and differences of Fritillaria anhuiensis Bulbus, providing reference for the quality evaluation of Fritillaria anhuiensis Bulbus and standardization of it.

AIM: To evaluate the bioequivalence of two kinds of amlodipine besylate tablets in Chinese healthy subjects under fasting and fed conditions. METHODS: Twenty-four healthy subjects were enrolled, and a random, open, single-dose, two preparations, two sequences and double-crossover design was used to give the test or reference preparations under fasting and postprandial conditions. The concentration of metformin in plasma was detected by LC-MS/MS, and the main pharmacokinetic parameters were calculated to evaluate the bioequivalence. RESULTS: In fasting state, the mean of C

Events including antibody‒antigen affinity, internalization, trafficking and lysosomal proteolysis combinatorially determine the efficiency of antibody-drug conjugate (ADC) catabolism and hence the toxicity. Nevertheless, an approach that conveniently identifies proteins requisite for payload release and the ensuing toxicity for mechanistic studies and quality assessment is lacking. Considering the plethora of ADC candidates under development, we developed a target-responsive subcellular catabolism (TARSC) approach that examines ADC catabolism and probes changes in response to targeted interferences of proteins of interest. We firstly applied TARSC to study the commercial T-DM1 and the biosimilar. We recorded unequivocal catabolic behaviors regardless of the absence and presence of the targeted interferences. Their negligible differences in TARSC profiles agreed with their undifferentiated anti-tumoral efficacy according to further

OBJECTIVE：To establish the m ethod for simultaneous determination of 8 kinds of ginsenosides in Panax quinquefolium broken pieces. METHODS ：HPLC-DAD method was used to determine the contents of ginsenoside Rg 1，Re，Rb1， Rc，Ro，Rb2，Rb3，Rd in P. quinquefolium broken pieces. The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of acetonitrile- 0.2％ phosphoric acid water solution （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃. The detection wavelength was set at 203 nm，and sample size was 10 μL. Ginsenoside Re and ginsenoside Rb 2 were used as control ，liner calibration with two-reference substances correction was used to predict the retention time of other 6 components，and was compared with the relative retention time method. Using ginsenoside Re as control ， above components were quantified by the relative correction factor method ，and the results were compared with the external standard method. RESULTS ：The contents of ginsenoside Rg 1，Re，Rb1，Rc，Ro，Rb2，Rb3，Rd were 10.59-12.78，2.160-2.768， 27.492-38.880，3.154-4.018，3.368-4.080，0.343-0.755，0.961-1.415，5.857-6.923 mg/g. The accuracy of two-reference substances linear correction method to predict the retention time of components was higher ，and the absolute deviation of the predicted retention time was lower than that of the relative retention time method. There was no significant difference between the relative correction factor method and the external standard method ，and relative error was ＜3％ . CONCLUSIONS ：Established two-reference substances for determination of multiple components can be used for qualitative and quantitative analysis of 8 kinds of ginsensides in P. quinquefolium broken pieces simultaneously and accurately.

OBJECTIVE：To establish the content determination me thod of 3 mono/disaccharides in 3 kinds of medicinal Dendrobii Caulis. METHODS ：HPLC-CAD method was established. The determination was performed on Shodex Asahipak NH2P-50 4E column with mobile phase consisted of acetonitrile-water （75 ∶ 25，V/V）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃，and sample size was 10 µL. CAD detection condition included that data acquisition frequency was 5 Hz，filter constant was 5 s，atomization temperature was 35 ℃ ，gas source was nitrogen with pressure of 4.012 × 105 Pa. RESULTS：The linear range of fructose ，D-anhydrous glucose and sucrose were 0.156 2-1.873 8 mg/mL（r＝0.999 5），0.012 7- 0.152 4 mg/mL（r＝0.999 7），0.277 6-3.331 2 mg/mL（r＝0.999 8），respectively. The limits of quantification were 0.002 61，0.004 24 and 0.005 12 mg/mL，and the limits of detection were 0.000 78，0.001 27 and 0.001 54 mg/mL，respectively. RSDs of precision ， stability，reproducibility and durability tests were all lower than 3％. The recoveries were 95.98％-98.15％（RSD＝0.83％，n＝6）， 95.64％-98.62％（RSD＝1.10％，n＝6）and 97.53％-98.94％（RSD＝0.53％，n＝6）. The contents of them were 0.28％-1.12％， 0.02％-0.13％，0.76％-2.67％，respectively. The total content was 1.38％～3.10％. The order of saccharide content in 3 kinds of Dendrobii Caulis was sucrose ＞fructose＞D-anhydrous glucose ；the order of sucrose content and total content were Dendrobium huoshanense＞D. moniliforme ＞D. officinale ；the order of D-anhydrous glucose content was D. huoshanense ＞D. officinale ＞D. moniliforme； the order of fructose content was D. moniliforme ＞D. officinale ＞D. huoshanense . CONCLUSIONS ：Established method is sensitive ，reproducible and simple in operation ，and can be used for content determination of 3 saccharides in 3 kinds of medicinal Dendrobii Caulis. There are differences in the contents of saccharide s among 3 kinds of Dendrobii Caulis.

Objective To investigate the influence of depression on glycemic control in the patients with type 2 diabetes mellitus(T2DM).Methods The Zung self-rating depression scale(SDS) was used to assess depression.A total of 276 cases of T2DM were divided into the group A(SDS standard score ≥53 points) and B(SDS standard score ＜53points).The levels of HbA1c,FPG,HOMA-IR,etc.were compared between the two groups,and the influencing factors of glycemic control in T2DM patients were analyzed.Results In the patients with T2DM,the SDS standard score was correlated with HbA1c(r=0.26,P＜0.05).The multivariate regression analysis showed that the SDS standard score was still correlated with HbA1c (β =0.30,t =5.1,P＜ 0.05).The HbA1c level in the group A was higher than that in the group B(t=3.685,P＜0.05);after correcting the factors of sex,age and education,the HbA1c level in the group A was still higher than that in the group B(F=47.8,P＜0.05).Conclusion The depression mood is adverse to glycemic control in T2DM patients.

Laccase is a widely-used environment-friendly copper-containing oxidase found in many plants, insects and fungi. Recently, more and more laccases are also found in bacteria. Myxobacteria are an important bacteria resource. However, myxobacteria are much more difficult to isolate and purify than other bacteria. We used bioinformatic approach to screen myxobacteria proteomes available in NCBI. Based on conserved sequences of four copper binding sites in multicopper oxidase, 30 potential laccase sequences were obtained. Among them, nine genes were synthesized and expressed in Escherichia coli BL21 (DE3). Seven proteins showed laccase activity when tested with traditional laccase substrates. One protein, named rSC-2, was chosen for further research because it exhibited the highest activity towards 2,6-dimethyl phenol (DMP). The molecular weight of rSC-2 was 57 kDa. Its specific activity to DMP was 0.27 U/mg. The optimal temperature and the optimal pH were 60 ℃ and 7.0, respectively. About 50% of the original activity was retained after incubation at 60 ℃ and pH 7.0-8.0 for 1 h. Metals showed different effects on rSC-2. rSC-2 activity was enhanced by several metalsat concentration of 1 mmol/L, such as Ca²⁺ and Mn²⁺. With a higher concentration of 5 mmol/L, the activity of rSC-2 was apparently inhibited. This is the first report of bioinformatics screening myxobacteria laccases in combination with expression in E. coli.