Effects of total ginsenosides from Panax ginseng stems and leaves on gut microbiota and short-chain fatty acids metabolism in acute lung injury mice / 中国中药杂志

This study aimed to investigate the biological effects and underlying mechanisms of the total ginsenosides from Panax ginseng stems and leaves on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in mice. Sixty male C57BL/6J mice were randomly divided into a control group, a model group, the total ginsenosides from P. ginseng stems and leaves normal administration group(61.65 mg·kg~(-1)), and low-, medium-, and high-dose total ginsenosides from P. ginseng stems and leaves groups(15.412 5, 30.825, and 61.65 mg·kg~(-1)). Mice were administered for seven continuous days before modeling. Twenty-four hours after modeling, mice were sacrificed to obtain lung tissues and calculate lung wet/dry ratio. The number of inflammatory cells in bronchoalveolar lavage fluid(BALF) was detected. The levels of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in BALF were detected. The mRNA expression levels of IL-1β, IL-6, and TNF-α, and the levels of myeloperoxidase(MPO), glutathione peroxidase(GSH-Px), superoxide dismutase(SOD), and malondialdehyde(MDA) in lung tissues were determined. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in lung tissues. The gut microbiota was detected by 16S rRNA sequencing, and gas chromatography-mass spectrometry(GC-MS) was applied to detect the content of short-chain fatty acids(SCFAs) in se-rum. The results showed that the total ginsenosides from P. ginseng stems and leaves could reduce lung index, lung wet/dry ratio, and lung damage in LPS-induced ALI mice, decrease the number of inflammatory cells and levels of inflammatory factors in BALF, inhibit the mRNA expression levels of inflammatory factors and levels of MPO and MDA in lung tissues, and potentiate the activity of GSH-Px and SOD in lung tissues. Furthermore, they could also reverse the gut microbiota disorder, restore the diversity of gut microbiota, increase the relative abundance of Lachnospiraceae and Muribaculaceae, decrease the relative abundance of Prevotellaceae, and enhance the content of SCFAs(acetic acid, propionic acid, and butyric acid) in serum. This study suggested that the total ginsenosides from P. ginseng stems and leaves could improve lung edema, inflammatory response, and oxidative stress in ALI mice by regulating gut microbiota and SCFAs metabolism.

Identification of key enzyme genes involved in biosynthesis of steroidal saponins and analysis of biosynthesis pathway in Polygonatum cyrtonema / 中国中药杂志

Steroidal saponins, which are the characteristic and main active constituents of Polygonatum, exhibit a broad range of pharmacological functions, such as regulating blood sugar, preventing cardiovascular and cerebrovascular diseases and anti-tumor. In this study, we performed RNA sequencing(RNA-Seq) analysis for the flowers, leaves, roots, and rhizomes of Polygonatum cyrtonema using the BGISEQ-500 platform to understand the biosynthesis pathway of steroidal saponins and study their key enzyme genes. The assembly of transcripts for four tissues generated 129 989 unigenes, of which 88 958 were mapped to several public databases for functional annotation, 22 813 unigenes were assigned to 53 subcategories and 64 877 unigenes were annotated to 136 pathways in KEGG database. Furthermore, 502 unigenes involved in the biosynthesis pathway of steroidal saponins were identified, of which 97 unigenes encoding 12 key enzymes. Cycloartenol synthase, the first key enzyme in the pathway of phytosterol biosynthesis, showed conserved catalytic domain and substrate binding domain based on sequence analysis and homology modeling. Differentially expressed genes(DEGs) were identified in rhizomes as compared to other tissues(flowers, leaves or roots).The 2 437 unigenes annotated by KEGG showed rhizome-specific expression, of which 35 unigenes involved in the biosynthesis of steroidal saponins. Our results greatly extend the public transcriptome dataset of Polygonatum and provide valuable information for the identification of candidate genes involved in the biosynthesis of steroidal saponins and other important secondary metabolites.

Efficacy and Material Basis of Shuangjin Sanjie Granules in Treatment on Hyperplasia of Mammary Gland / 中国实验方剂学杂志

Objective:The SD rat model of hyperplasia of mammary gland（HMG） and the ultrahigh-performance liquid chromatography and mass spectrometry (UHPLC-LTQ-Orbitrap MS) technology were used to explore the pharmacological material basis of Shuangjin Sanjie granules (SJSJG) for the treatment on HMG.Method:SD rat models of HMG were administered in groups, and the nipple height and the diameter were measured; the levels of estradiol (E2), progesterone (P) and prolactin (PRL) in serum were detected, pathological examination was conducted for the hyperplasia of breast tissue. Histochemical methods were used to detect the expressions of estrogen receptor α (ERα), androgen receptor (AR), progesterone receptor (PR), tumor necrosis factor-α (TNF-α) proteins. Finally, UHPLC-LTQ-Orbitrap MS technology was used to detect the main chemical constituents of SJSJG, and the pharmacodynamic substance basis was analyzed based on the pharmacological effect.Result:The results of animal experiments showed that compared with the normal group, nipple height and diameter of the model group increased remarkably (P<0.01), serum E2 significantly increased (P<0.01). Pathological examination showed abnormal hyperplasia of breast tissue， expressions of ERα, AR, PR and TNF-α increased， compared with the model group, the nipple height and diameter of the SJSJG group decreased remarkably (P<0.01), serum E2 was decreased significantly (P<0.01)， pathological examination showed weakened abnormal hyperplasia of breast tissue, ERα, AR, PR and TNF-α protein expressions were significant decreased (P<0.01). The results of basic material study showed that 85 chemical components were identified from SJSJG, including 16 alkaloids, 7 flavonoids, 15 terpenes, 9 phenolic acid compounds, 3 coumarin compounds, 10 esters and lactone compounds, 7 fatty acids compounds, 4 amino acids compounds, and 14 other types of ingredients， among them, alkaloids and terpenoids chemical drug substances were closely related.Conclusion:SJSJG can effectively improve the condition of breast hyperplasia, and its medicinal substance basis may include saikosaponin A, Saikosaponin D, verticinone, peimine.

Literature Research on Traditional Chinese Medicine Therapeutic Principles and Methods and Common Prescriptions of Benign Prostatic Hyperplasia / 中国实验方剂学杂志

Objective:Define traditional Chinese medicine (TCM) therapeutic principles and methods and common prescriptions of benign prostatic hyperplasia (BPH) based on clinical experience of famous doctors. Method:China National Knowledge Infrastructure, China Biology Medicine disc, China Science and Technology Journal Database, Wanfang database were systematically retrieved for literatures of famous doctors in diagnosis and treatment of BPH from January 1, 1997 to December 31, 2018.A literature analysis database was established for integration and analysis of relevant content of TCM therapeutic principles and methods and prescriptions. SPSS 20.0 software was used for statistics to obtain TCM therapeutic principles and methods, common prescriptions and drug distribution regularity of BPH. Result:A total of 109 qualified literatures were included in this study, involving 66 famous doctors and experts, and 9 kinds of BPH therapeutic principles were obtained, including "treatment of both symptoms and root cause of disease", "tonification and purgation in combination", "coordination of Yin and Yang". There are 55 kinds of therapies, including "promoting blood circulation for removing blood stasis", "clearing heat and promoting dampness", "warming and recuperating kidney Yang", "softening and resolving hard mass" and " invigorating kidney Qi". And BPH was mostly diagnosed and treated based on kidney, spleen and lung. Through collection and summarization, totally 38 formulas commonly used in treating BPH obtained. The most commonly used ones were "Guizhi Fuling Wan", "Zishen Tongguan Wan", "Buzhong Yiqi Tang", " Bazheng Tang" and "Jisheng Shenqi Wan". There were 217 commonly used herbs, mainly including " Astragali Radix", "Rehmanniae Radix Praeparata", "Atractylodis Macrocephalae Rhizoma", "Glycyrrhizae Radix et Rhizoma", "Angelicae Sinensis Radix", "Radix Achyranthis Bidentatae", "Peach Kernel", "Pangolin Scales", "Vaccariae Semen", "Salviae Miltiorrhizae Radix et Rhizoma", "Poria", "Rhizoma Alismatis" and "Plantain Seed". Conclusion:The treatment of BPH is based on the principles of "treatment of both symptoms and root cause of disease, and tonification and purgation in combination". The commonly used therapies include prescription for "promoting blood circulation for removing blood stasis", "clearing heat and promoting dampness" and "warming and recuperating kidney Yang". The corresponding prescriptions shall be based on symptoms.

Cloning and characterization of chalcone synthase and chalcone isomerase genes in Arisaema heterophyllum / 中国中药杂志

Chalcone synthase( CHS) and chalcone isomerase( CHI) are key enzymes in the biosynthesis pathway of flavonoids. In this study,unigenes for CHS and CHI were screened from the transcriptome database of Arisaema heterophyllum. The open reading frame( ORFs) of chalcone synthase( Ah CHS) and chalcone isomerase( Ah CHI) were cloned from the plant by RT-PCR. The physicochemical properties,expression and structure characteristics of the encoded proteins Ah CHS and Ah CHI were analyzed. The ORFs of Ah CHS and Ah CHI were 1 176,630 bp in length and encoded 392,209 amino acids,respectively. Ah CHS functioned as a symmetric homodimer. The N-terminal helix of one monomer entwined with the corresponding helix of another monomer. Each CHS monomer consisted of two structural domains. In particular,four conserved residues define the active site. The tertiary structure of Ah CHI revealed a novel open-faced β-sandwich fold. A large β-sheet( β4-β11) and a layer of α-helices( α1-α7) comprised the core structure. The residues spanning β4,β5,α4,and α6 in the three-dimensional structure were conserved among CHIs from different species. Notably,these structural elements formed the active site on the protein surface,and the topology of the active-site cleft defined the stereochemistry of the cyclization reaction. The homology comparison showed that Ah CHS had the highest similarity to the CHS of Anthurium andraeanum,while Ah CHI had the highest similarity to the CHI of Paeonia delavayi. This study provided the basis for the functional study of Ah CHS and Ah CHI and the further study on plant flavonoid biosynthesis pathway.

Cloning and heterologous expression of sansanmycin biosynthetic gene cluster in Streptomyces coelicolor / 药学学报

Sansanmycins (SSs), produced by Streptomyces sp. SS, belong to uridyl peptide antibiotics which exhibit a good inhibitory effect on Mycobacterium tuberculosis and Pseudomonas aeruginosa. They share a unique chemical scaffold with a 4',5'-enamide-3'-deoxyuridine attached to DABA (N-methyl-2,3-diaminobutyryl) which was located in the peptide chain through peptide bond. In order to study the function of related genes and to employ synthetic biology to gain new SS derivatives, we obtained a complete SS biosynthetic gene cluster and heterologously expressed it in Streptomyces coelicolor M1146, M1152 and M1154. Fermentation broth of the recombinant strains were detected using HPLC and HPLC-MS/MS, and the result showed that SS-A was successfully produced in the three strains, and its production level in S. coelicolor M1154 was similar to the original wild type strain. In addition, a potential SS analogue named as SS-1154 was discovered from the fermentation broth of S. coelicolor M1154.

The Effect of Serum Cytokine Levels prior Transplantation on the Outcome of Severe Aplastic Anemia Patients Received Allogeneic Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To identify the role of serum cytokine levels prior allogeneic hematopoietic stem cell trans-plantation (allo-HSCT) in the outcome of severe aplastic anemia (SAA) patients received allo-HSCT treatment.</p><p><b>METHODS</b>The clinical data of 117 SAA patients received allo-HSCT were enrolled in this study. The overall survival (OS), graft versus host disease (GVHD) incidence and relationship of serum cytokines with OS and major transplantation complications were retrospectively analyzed.</p><p><b>RESULTS</b>The patients enrolled in this study included 78(66.7%) cases received HSCT matched sibling donors (MSD), 12(10.2%) HSCT of unrelated donors (MUD) and 27 cases received HSCT of haploidentical donors (HID). The 5-years OS was 76.0%(95% CI: 64.4-87.5%); aGVHD cumulative incidence was 49.6%(95% CI: 40.4%-58.8%) and cumulative incidence cGVHD was 31.6%(95% CI:23.1%-40.2%). MSD allo-HSCT had a significantly higher 5-years OS as compared with the other donors(82.3%±6.6% vs 61.3%±11.7%, P<0.05). HLA matching, donor's age, cytomegalovirus/ Epstein-Barr virus (CMV/EBV) infection were important factors of affecting occurence of aGVHD. The patients with higher serum IL-6 had reduced platelet recovery time after transplantation (14.6±1.8 vs 18.3±2.6 d)(P=0.050) and higher serum TNF-α level accompanied by a lower incidence of CMV/EBV infection (37.8%±11.1% vs 58.8±16.8%)(P<0.05).</p><p><b>CONCLUSION</b>MSD allo-HSCT is the effective treatment for SAA patients. Donor's type remains the strong predictor of survival. The serum levels IL-6 and TNF-α before transplantation associate with platelet recovery and CMV/EBV infection.</p>

Chemical profile of the active fraction of Yi-Gan San by HPLC-DAD-Q-TOF-MS and its neuroprotective effect against glutamate-induced cytotoxicity / 中国天然药物

Yi-Gan San (YGS), a traditional Chinese medicine for dementia-related symptoms, was previously fractionated. One active fraction, YGS40 exhibited a neuroprotective effect against glutamate-induced cytotoxicity. In the present study, high-performance liquid chromatography, coupled with diode-array detection and quadrupole time-of-flight mass spectrometry, was applied for the identification of its chemical constituents and for quantification studies. The chemical profile of YGS40 consisted of sixty-four identified or tentatively characterized compounds. The levels of the major marker compounds increased significantly in the mixed decoction compared with those in the single plant decoction. The results suggest the high precision of the analyses of most of the constituents in YGS40 and establish the quantitative variations of the major marker compounds between the single and mixed decoction processes.

Alcohol-induced proliferation of neurons in mouse hippocampal dentate gyrus: a possible role of ceramide / 生理学报

To investigate the role and mechanism of ceramide (Cer) regulation in alcohol-induced neuronal proliferation and the newborn neurons formation, we used sphingomyelin synthase 2 (predominant enzyme of Cer metabolism) knockout (SMS2(-/-)) and wild type (WT) female mice to establish the model of prenatal alcohol exposure. In 24 h after being given birth (postnatal day 0, P0), the offspring of model mice received blood sphingomyelin (SM) measurement with enzymatic method. On P0, P7, P14 and P30, the proliferation of granule cells in the dentate gyrus and newborn neurons were investigated with immunofluorescent labeling. The expression of protein kinase Cα (PKCα) in the hippocampus was tested with Western blot analysis. The results showed that the SM level of blood in SMS2(-/-) pups was significantly lower than that in WT pups. No matter in SMS2(-/-) or WT mice, the prenatal alcohol exposure down-regulated the SM levels in pups with dose-dependency. In both SMS2(-/-) and WT pups, the number of proliferative neurons and newborn neurons in the dentate gyrus gradually decreased with the growing age. Compared with the WT pups, SMS2(-/-) pups showed significantly more proliferative neurons and newborn neurons in the dentate gyrus. Notably, prenatal alcohol exposure dose-dependently increased proliferative neurons and newborn neurons in the dentate gyrus in both WT and SMS2(-/-) pups. The hippocampal expression of PKCα protein in SMS2(-/-) mice was lower than that in WT mice, and prenatal alcohol exposure could up-regulate the PKCα protein expression in both WT and SMS2(-/-) mice with dose dependency. These results suggest that alcohol exposure during pregnancy can induce the compensatory neural cell proliferation and the production of newborn neurons in offspring, and the Cer-ceramide-1-phosphate (C1P) pathway is involved in alcohol-induced neural cell proliferation. The activation of PKCα may be a key step to start the Cer-C1P pathway and up-regulate the alcohol-induced neural cell proliferation and the newborn neurons formation.

Sphingomyelin synthase 2 deficiency decreases atherosclerosis and inhibits inflammation in mice / 生理学报

Plasma sphingomyelin (SM) has been shown to be an independent risk factor for coronary heart disease, and sphingomyelin synthase 2 (SMS2) contributes to de novo SM biosynthesis and plasma membrane SM levels. The aim of the present study is to evaluate the in vivo role of SMS2 deficiency in serum SM metabolism and atherosclerosis (AS) development. We used male SMS2 knockout (SMS2(-/-)) and C57BL/6J (wild-type, WT) mice as experimental and control groups, respectively. Each group was fed high-fat diet (1% cholesterol, 20% leaf fat), as well as bile salt for accelerating the atherosclerotic formation. After three months of feeding, the mice were killed to observe aortic arches and oil red-stained longitudinal sections of thoracoabdominal aortae. Fasting blood samples were taken from the tail vein before and after high-fat diet, and the serum lipid and SM levels were measured by using kits and enzymatic method respectively. Western blot was used to analyze the contents of nuclear factor-kappaB (NFkappaB) p65 subunit in peritoneal macrophages stimulated with lipopolysaccharide (LPS) after high-fat diet. The results showed that after high-fat diet, SMS2(-/-) mice presented decreased atherosclerotic lesions in aortic arch and thoracoabdominal aorta compared with WT mice. Regardless of whether high-fat diet were given or not, SMS2(-/-) mice showed a significant decrease in serum SM level (P<0.05), but no significant changes in serum lipid levels, compared with WT mice. The expressions of NFkappaB p65 were attenuated in macrophages from SMS2(-/-) mice in response to LPS stimulation compared with those of the WT mice. These results suggest that SMS2 deficiency decreases AS and inhibits inflammation in mice. Thus, SMS2 deficiency may be a potential therapeutic strategy.

Quinovic acid triterpenoid saponins from bark of Mitragyna rotundifolia / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from the bark of Mitragyna rotundifolia.</p><p><b>METHOD</b>Column chromatographic techniques were applied to isolate constituents. A combination of IR, MS and NMR spectroscopy was used to identify structures of constituents.</p><p><b>RESULT</b>Six compounds were isolated from the n-BuOH fraction and their structures were elucidated as quinovic acid-3-O-beta-D-6-deoxy-glucopyranoside, 28-O-beta-D-glucopyranosyl ester (1), quinovic acid-27-O-alpha-L-rhamnopyranosyl ester (2), quinovic acid-3-O-alpha-L-rhamnopyranoside (3), qunovic acid-27-O-beta-D-glucopyranosyl ester (4), quovic acid-3-O-beta-D-6-deoxy-glucopyranoside (5), qunovic acid-27-O-beta-6-deoxy-D-glucopyranosyl ester (6).</p><p><b>CONCLUSION</b>Compounds 1 - 6 were isolated for the first time from the plant. Compounds 1 - 4 and 6 were isolated for the first time form the genus.</p>