Integrating chemical similarity and bioequivalence: an overall evaluation of the quality consistency of traditional decoction and dispensing granule decoction of Amomum villosum.

OBJECTIVE: This study aims to investigate the quality consistency between traditional decoction (TD) of Amomum villosum and its dispensing granule decoction (DGD). Fifteen batches of TD and nine batches of dispensing granules (manufactured by A, B, and C) were prepared and evaluated for their consistency. METHODS: Firstly, The chemical similarity of TD and DGD was examined using GC and HPLC, coupled with hierarchical cluster analysis (HCA), criteria importance though intercrieria correlation(CRITIC) weighting method, and principal component analysis (PCA). Secondly, the gastrointestinal motility experiments in mice, along with the CRITIC weighting method, were employed to assess the bioequivalence of TD and DGD of Amomum villosum. Finally, the entropy weight technique-gray relative analysis(GRA) method was used to compare the quality of Amomum villosum decoctions. RESULTS: ①The CRITIC weighting method indicated significantly higher scores for TD than DGD (p < 0.01). HCA and PCA results demonstrated a clear distinction between TD and DGD. ②Gastrointestinal motility test results revealed no significant difference between TD and DGD in other indicators (p > 0.05).③Gray relative analysis results showed that the relative correlation of TD was more significant than that of DGD. CONCLUSION: The chemical composition of DGD and TD differed. The biological activity of DGD-A/B was consistent with that of TD, while the difference between DGD-C and TD was significant. A comprehensive evaluation showed that TD exhibited better quality than DGD. DGD manufacturers should optimize the preparation process to enhance product quality.

[Main characteristics and practical discussions of national drug standards for 196 kinds of Chinese medicine formula granules].

As China is implementing the policy of "Announcement on Ending the Pilot Work of Chinese medicine formula gra-nules", the standard of Chinese medicine formula granules has gradually become the focus of industry development. Up to now, 196 national drug standards for Chinese medicine formula granules have been published by China, which guaranteed the production quality of Chinese medicine formula granules. However, there are still several challenges such as the rational application of national drug standards and the enrichment and improvement of varieties. The basic content of the issued national drug standards for Chinese medicine formula granules was analyzed and compared with the quality standard provisions of the corresponding decoction pieces in the Chinese Pharmacopoeia(2020 edition) in this paper. This paper discussed the main characteristics of paste-forming rate of each medicinal raw materials, "quantity-quality" transformation, equivalent ratio, and so on, and clarified the characteristics of the national standard for Chinese medicine formula granules. This paper provided references for achieving the unified quality control and meeting the overall quality requirements of Chinese medicine formula granules.

A comparative study of consistency between Paeonia rubra hort traditional decoction and dispensing granule decoctions from different sources.

OBJECTIVE: To compare the consistency between the decoction of Paeonia rubra hort dispensing granules from different manufacturers and traditional decoction (TD), and to provide a reference for the clinical application of Paeonia rubra hort dispensing granules. METHODS: Nine batches of Paeonia rubra hort dispensing granules (from three manufacturers, A, B, and C) and 20 batches of Paeonia rubra hort decoction pieces were collected. The contents of four active components in vivo and in vitro were determined by HPLC and UPLC-MS/MS, respectively. The consistency of the Paeonia rubra hort decoction pieces and dispensing granules were compared based on the Criteria Importance Though Intercrieria Correlation (CRITIC) weighting method and the equivalent correction suggestions (1 g of dispensing granule equals the same amount of Chinese herbal samples) were put forward for the dispensing granules. RESULTS: The total content of active ingredients in vivo and in vitro of manufacturer A was significantly lower than that of TD (p < 0.05), and the total content of active ingredients in vivo of manufacturer C was significantly lower than that of TD (p < 0.05); The equivalent of manufacturer A and manufacturer C should be corrected from 1:11 and 1:5 to 1:5 and 1:4, respectively. CONCLUSION: The quality of Paeonia rubra hort dispensing granule decoction from some manufacturers aligns that of TD, but the other is slightly inferior to that of TD. After appropriate equivalent correction, quality consistency can be achieved with TD.

In vitro facilitating role of polygonatum sibiricum polysaccharide in osteogenic differentiation of bone marrow mesenchymal stem cells from patients with multiple myeloma.

BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) were proved to play a vital role in multiple myeloma (MM). Polygonatum sibiricum polysaccharide (PSP) was found to have anti-tumor pharmacological effects, yet its interaction with BMMSCs remained poorly understood. Therefore, we explore the effect of PSP on osteogenic differentiation of BMMSCs. METHODS: BMMSCs were isolated by density gradient centrifugation. CD90 and CD34 were detected by flow cytometry (FCM). Osteogenic marks were detected by quantitative real-time PCR (qRT-PCR) and Western blotting (WB). The vitality of cells treated with different concentrations of PSP was observed by Cell Counting Kit-8 (CCK-8). ALP staining kit was used to detect the activity of alkaline phosphatase (ALP). Alizarin red staining detected the formation of mineralized nodules. Osteoblast-associated genes were evaluated by qRT-PCR and WB. The phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) signaling pathways were tested by WB. RESULTS: The BMMSCs showed good growth under an inverted microscope. FCM showed that CD34 and CD45 was low-expressed, whereas CD44, CD90 and CD105 was highly expressed. Compared with the Control group, the expressions of Runx2 and ALP in cells were significantly increased. CCK-8 showed that different concentrations of PSP had no significant effect on the viability of BMMSCs. BMMSCs treated with 25 mg/l PSP were stained the most deeply by ALP. Mineralized nodules in PSP groups dramatically increased, and hit a peak under the action of 25 mg/l PSP. PSP up-regulated p-PI3K, p-AKT, and p-mTOR, but had no significant effect on PI3K, AKT, and mTOR. CONCLUSION: PSP induced osteogenic differentiation of BMMSCs from MM patients.

Study on the effect of Mongolian medicine Qiwei Qinggan Powder on hepatic fibrosis through JAK2/STAT3 pathway.

This research aimed to evaluate the antihepatic fibrosis effect and explore the mechanism of Qiwei Qinggan Powder (QGS-7) in vivo and in vitro. Carbon tetrachloride (CCl4)-treated rats and hepatic stellate cells (HSCs) were used. QGS-7 treatment significantly improved the liver function of rats as indicated by decreased serum enzymatic activities of alanine aminotransferase, aspartate transaminase, and alkaline phosphatase. Meanwhile, the hydroxyproline of liver was significantly decreased. Histopathological results indicated that QGS-7 alleviated liver damage and reduced the formation of fibrosis septa. Moreover, QGS-7 significantly attenuated expressions of Alpha smooth muscle actin, Collagen I, Janus kinase 2 (JAK2), phosphorylation-JAK2, signal transducer and activator of transcription 3 (STAT3), phosphorylation-STAT3 in the rat hepatic fibrosis model. QGS-7 inhibited HSC proliferation and promoted it apoptosis. QGS-7 may affect hepatic fibrosis through JAK2/STAT3 signaling pathway so as to play an antihepatic fibrosis role.

[Investigation of benzo(a)pyrene contamination in edible vegetable oil sold in Henan Province from 2017 to 2019].

OBJECTIVE: To investigate the contamination of benzo(a)pyrene in edible vegetable oil from Henan Province. METHODS: A total of 1190 edible vegetable oil samples from Henan Province were collected. The concentration of benzo(a)pyrene was examined by high performance liquid chromatography. RESULTS: The concentration of benzo(a)pyrene in prepackaged edible vegetable oils was <0. 3-63. 5 µg/kg with an average of 1. 5 µg/kg. The detection rate of benzo(a)pyrene in samples was 80. 0%, and the over standard rate was 0. 6%. In terms of subclasses of vegetable oil, the sesame seed oil samples were most seriously contaminated with benzo(a)pyrene. The risk of the benzo(a)pyrene contamination of sesame seed oil produced by aqueous extraction method was higher than that by mechanical pressed method. The risk of benzo(a)pyrene contamination in sesame seed oil and rapeseed oil increased as the decrease of quality grade. CONCLUSION: The result suggested that the sesame seed oil produced by aqueous extraction method had high risk in benzo(a)pyrene contamination in Henan Province.

Anti-liver fibrosis effect of total flavonoids from Scabiosa comosa Fisch. ex Roem. et Schult. on liver fibrosis in rat models and its proteomics analysis.

BACKGROUND: To explore the potential therapeutic effect of total flavonoids (TFs) extracted from Scabiosa comosa Fisch. ex Roem. et Schult on liver fibrosis in rat models and to identify the possible targets and pathways of TF in treating liver fibrosis by using a quantitative proteomics method. METHODS: Sixty Wistar rats were equally randomized into five groups: a blank control group, a model group, and high-, intermediate-, and low-dose TF treatment groups. Except for the blank control group, rats in the other four groups were intragastrically administered with CCL4 2 mL/kg to establish the liver fibrosis models. Furthermore, the high-, intermediate-, and low-dose TF groups were intragastrically given TF at a dose of 200, 100 and 50 mg/kg, respectively. After 10 weeks, the rats were sacrificed, and blood and liver samples were collected. Serum alanine transaminase (ALT), Aspartate aminotransferase (AST), and alkaline phosphatase (ALP) levels were measured, and hematoxylin and eosin (HE) staining and Masson's trichrome staining were used to observe the pathological changes in each group. The hydroxyproline content was also determined. Real-time polymerase chain reaction (PCR) and Western blotting (WB) were performed to detect the mRNA and protein expressions of α-smooth muscle actin (αSMA) and Collagen I. Mass spectrometry was performed for proteomic analysis. RESULTS: Compared with the blank control group, the model group had significantly higher ALT, AST, ALP, and hydroxyproline levels; also, HE and Masson staining showed fibrotic lesions and inflammatory cell infiltration in the model group. Compared with the model group, the high-, intermediate-, and lowdose TF groups had significantly decreased ALT, AST, and ALP levels (P<0.05), and a significantly lower hydroxyproline level (P<0.05), along with remarkably improved fibrotic lesions and inflammatory cell infiltration. Real-time PCR and WB showed that the model group had significantly higher expressions of αSMA and collagen I than those in the blank control group, whereas the TF groups had significantly lower expressions of αSMA and collagen I than those in the model group. A total of 5,014 proteins were detected by quantitative proteomics, among which 205 proteins were differentially expressed, 77 of which were upregulated and 128 of which were down-regulated. KEGG pathway analysis indicated that the peroxisome proliferator activated receptor (PPAR) and ECM-receptor interaction pathways were down-regulated in the TF groups compared with the model group. Among them, fatty-acid-binding protein (FABP) and von Willebrand factor (vWF) were the key proteins in the PPAR and extracellular matrix (ECM)-receptor interaction pathways. The proteomic results were validated by using WB, yielding consistent results. CONCLUSIONS: Our result demonstrated that the TF extract of Scabiosa comosa Fisch. ex Roem. et Schult has a good anti-liver fibrosis effect and may prevent liver fibrosis by reducing the content of α-SMA, CollagenⅠ in liver tissue. The anti-fibrosis mechanism of TF extract of Scabiosa comosa Fisch. ex Roem. et Schult may be the inhibition of key proteins FABP and vWF in PPAR, ECM RECEPTOR INTERACTION pathway.

Effects of parathyroidectomy on bone metabolism in haemodialysis patients with secondary hyperparathyroidism.

OBJECTIVE: To evaluate the outcome of bone metabolism and bone mineral density (BMD) in haemodialysis patients after parathyroidectomy (PTX). METHODS: A total of 31 haemodialysis patients with secondary hyperparathyroidism (SHPT) were treated with PTX. BMD of lumbar spine (LS) and femoral neck (FN) was determined by dual energy X-ray absorptiometry. RESULTS: Parathyroidectomy ledds to significant decrease of serum ß-crosslaps (ß-CTX), osteocalcin (OC) and procollagen type I amino-terminal propeptide (PINP) while serum sclerostin (SOST) increased after surgery. BMD was markedly improved in both LS and FN after PTX. Z-scores analysis further confirmed that PTX significantly benefited bone metabolism in haemodialysis patients, which well correlated with the improvement of serum iPTH and OC. CONCLUSIONS: Parathyroidectomy leads to significant improvement of serum OC, PINP, ß-CTX and SOST, which may beneficially modify calcium-phosphorus metabolism and BMD in haemodialysis patients with SHPT.

[Effect of Cynomorium songaricum polysaccharide on telomere of lung cancer A549 cells].

To study the effect of Cynomorium songaricum polysaccharide (CSRP) on A549 cells telomere of human non-small cell lung cancer, the mice were intragastric administrated with CSRP (0.08 g•kg⁻¹) once daily for 4 days. Then their serum was taken for preparing CSRP drug serum. A549 cells were treated by the drug serum, and the effect of drug serum with different concentrations and different treating time on the proliferation of non-small cell lung cancer A549 cells was determined by MTT test. After treating for 48 hours by the drug serum of different concentrations, the telomere length of the cells was determined by fluorescence quantitative polymerase chain reaction (qPCR); the mRNA expression of telomerase reverse transcriptase (TERT) was determined by RT-qPCR; the cells apoptosis was determined by TUNEL assay. The results demonstrated that CSRP of various concentrations could inhibit the proliferation of the lung cancer A549 cells significantly, and the inhibition effect was strongest at 48 hours with the concentration of 6.0 mL•L⁻¹. At 48 h, that CSRP of the concentrations from 1.5 to 12.0 mL•L⁻¹ could significantly shorten the telomere length of A549 cells, and the effect was strongest with the concentration of 1.5 mg•L⁻¹. CSRP of various concentrations could significantly inhibit the mRNA expression of TERT in A549 cells, and the inhibition effect was stronger when the concentration was ≥6.0 mL•L⁻¹. CSRP of various concentrations could promote A549 cells apoptosis, and the effect was stronger when the concentration was ≥6.0 mL•L⁻¹. In conclusion, CSRP has the anti-cancer effect, and the action mechanism may be associated with inhibiting TERT mRNA expression, shortening telomere length, inhibiting cells proliferation and promoting cells apoptosis.

[Determination of prim-O-glucosylcimifugin and 5-O-methylvisammisoide in Saposhnikovia divaricata and HPLC fingerprint analysis].

OBJECTIVE: To develop HPLC methods for the determination of prim-O-glucosylcimifugin and 5-O-methylvisammisoide in Saposhnicovia divaricata and of HPLC fingerprint to compare the wild and culture varieties. METHOD: Conditions of determination: Shimadzu C18 column (4.6 mm x 150 mm, 5 microm), methanol-water (40:60) as mobile phase with the flow rate of 1 mL x min(-1). The detection wavelength was 254 nm. Conditions of HPLC fingerprint: MG II C18 column (4.6 mm x 250 mm, 5 microm), the mobile phase was acetonitrile-water with the flow rate of 1 mL x min(-1), using linear gradient elution, the column temperature was 30 degrees C. RESULT: The average recovery of prim-O-glucosylcimifugin was 99.6% (RSD 0.72%, n=6). The average recovery of 5-O-methylvisammisoide was 102.6% (RSD 0.88%, n=6). The contents of prim-o-glucosylcimifugin in wild and culture varieties were (4.96 +/- 2.59) and (3.61 +/- 1.82) mg x g(-1) respectively. The contents of 5-O-methylvisammisoide were (3.91 +/- 2.09) and (4.37 +/- 2.02) mg x g(-1) respectively. The compositions of S. divaricata were effective separated under the conditions of HPLC fingerprint. CONCLUSION: The HPLC determination method of prim-O-glucosylcimifugin and 5-O-methylvisammisoide is convenient and accurate. The HPLC fingerprint analysis method could be a basis for quality control and classification evaluate of S. divaricata.

[Effects of high dose methylprednisolone on cell apoptosis and Bcl-2 expression after acute spinal cord injuries in rats].

OBJECTIVE: To investigate the effect of high dose methylprednisolone (MP) on cell apoptosis and Bcl-2 expression after acute spinal cord injury (ASCI). METHODS: The 48 female rats were randomly divided into two groups:control group and experimental group. The spinal cord of rats were wounded at the level of T9,10 in moderate degree for each group. Thirty minutes after ASCI, the subjects in experimental group received MP injection through intraperitoneal with dosage of 30 mg/kg. Then a dosage of 5.4 mg/kg of MP was given through intraperitoneal injection every hour in 24 hours. The subjects in the control group received Normal Saline instead of MP with the same method. The impaired spinal cords were harvested on 4 h, 8 h, 1 d, 3 d, 7 d and 14 d after injury,and at each time point 4 rats were sacrificed in each group. The terminal deoxynucleotidal transferase-mediated DUTP-biotin nick end labeling (TUNEL) and immunohistochemical measurement were used to observe neural apoptosis and Bcl-2 expression. RESULTS: Apoptosis cells were noted primarily at 4 h after injury, with a maximum presence in lesion center at 8 h. The apoptotic index of MP-treated group had distinctly less than that of control group in 8 h, 1 d and 3 d after trauma. The Bcl-2 increased in the experimental group. CONCLUSION: The administration of the high dose methylprednisolone (MP) can inhibite the apoptosis after acute spinal cord injury in rats and increase the expression of Bcl-2.

[Effect of Cynomorium songaricum polysaccharide on telomere length in blood and brain of D-galactose-induced senescence mice].

OBJECTIVE: To study the effects of Cynomorium songaricum polysaccharide (CSP) on telomere length in blood and brain tissues of aged mice in order to provide some evidence for CSP's development and applying in the clinical uses. METHOD: Kunming mice were intraperitoneal injected D-galactose (500 mg x kg(-1) x d(-1)) to make the aging models, and different dosages of CSP (20, 40, 80 mg x kg(-1)) were given by gavage for 56 days. The average length of telomere was determined by real-time fluorescence quantitative PCR. RESULT: The relative T/S ratio of the group high and middle dosages of CSP in blood were 1.64 +/- 0.36 and 1.33 +/0.28, respectively, and higher than that of the group of senescence 1.01 +/- 0.13 (P < 0.01). Values of the group of high, middle, and low dosages of CSP in brain tissues were 3.34 +/- 0.58, 2.30 +/- 0. 75 and 1.55 +/- 0.58, respectively, and significantly higher than that of the group of senescence 1.04 +/- 0.33 (P < 0.01). CONCLUSION: CSP can exert the anti-aging effects by increase telomere length f senescence mice.