Anti-hyperuricemia activity and its mechanism of flavonoid extract from saffron floral bio-residues / 中国中药杂志

A hyperuricemic rat model induced by adenine and ethambutol was established to investigate the anti-hyperuricemia activity and its mechanism of the flavonoid extract from saffron floral bio-residues. Sixty-seven SD rats were randomly divided into control group, model group, positive control group, and flavonoid extract groups(with 3 doses), respectively, and each group contained 11 or 12 rats. The hyperuricemic model was established by continuous oral administration of adenine(100 mg·kg~(-1)) and ethambutol(250 mg·kg~(-1)) for 7 days. At the same time, the positive control group was given allopurinol(20 mg·kg~(-1) per day) and the flavonoid extract groups were given the flavonoid extract at doses of 340, 170 and 85 mg·kg~(-1) per day, respectively. On day 8, rat serum, liver, kidney, and intestinal tissues were collected, and the levels of uric acid in serum and tissue, the xanthine oxidase activities and antioxi-dant activities in serum and liver were evaluated, and the kidney histopathology was explored. In addition, an untargeted serum metabolomics study was performed. According to the results, the flavonoid extract effectively reduced the uric acid levels in serum, kidney and ileum and inhibited the xanthine oxidase activities and elevated the antioxidant activities of serum and liver in hyperuricemic rat. At the same time, it reduced the levels of inflammation factors in kidney and protected renal function. Moreover, 68 differential metabolites of hyperuricemic rats were screened and most of which were lipids and amino acids. The flavonoid extract significantly retrieved the levels of differential metabolites in hyperuricemic rats, such as SM(d18:1/20:0), PC[18:0/18:2(92,12Z)], palmitic acid and citrulline, possibly through the following three pathways, i.e., arginine biosynthesis, glycine, serine and threonine metabolism, and histidine metabolism. To sum up, the flavonoid extract of saffron floral bio-residues lowered the uric acid level, increased the antioxidant activity, and alleviated inflammatory symptoms of hyperuricemic rats, which may be related to its inhibition of xanthine oxidase activity and regulation of serum lipids and amino acids metabolism.

Risk factors of newly developed nonalcoholic fatty liver disease after pancreaticoduodenectomy / 中华外科杂志

Objective: To identify the risk factors of newly developed nonalcoholic fatty liver disease(NAFLD) after pancreaticoduodenectomy(PD). Methods: The clinical data of 130 patients who had undergone PD at Department of Hepatopancreatobiliary Surgery, Nanjing Drum Tower Hospital, the Affiliated Hospital of Medical School of Nanjing University from June 2018 to December 2020 were collected retrospectively. There were 74 males and 56 females, with age(M(IQR)) of 62(16) years (range: 22 to 84 years). Twenty-nine patients who developed NAFLD were divided into NAFLD group and 101 patients who did not suffer NAFLD were divided into no NAFLD group. Observation indications included:(1)preoperative demographics,intraoperative and postoperative characteristics; (2)the risk factors of newly developed NAFLD after PD. Count data were analyzed using χ2 test or Fisher's exact test. Measurement data were analyzed by student t test or Mann-Whitney U test. Multivariate analysis was performed using Logistic regression model with a stepwise forward approach. Results: All 130 patients successfully underwent PD and 29 cases(22.3%) developed NAFLD in 6 months after PD. The results of univariate analysis showed that gender,diabetic mellitus,the level of triglyceride preoperatively,and pancreatic ductal adenocarcinoma were the related factors of the development of NAFLD after PD(t=-2.655, χ²=4.563,U=-2.192,χ²=7.044;all P<0.05).Multivariate analysis revealed that gender,body mass index and pancreatic ductal adenocarcinoma were independent risk factors for the development of NAFLD after PD(OR=2.849,1.214,4.165,all P<0.05). Conclusion: Gender, body mass index and pancreatic ductal adenocarcinoma were independent risk factors for the development of NAFLD after PD.

Attenuation Mechanism of Aconti Lateralis Radix Praeparata Combined with Glycyrrhizae Radix et Rhizoma Based on Heart CYP450 Isozymes / 中国实验方剂学杂志

<b>Objective::To investigate the effect of Aconiti Lateralis Praeparata Radix combined with Glycyrrhizae Radix et Rhizoma on mRNA expression of cytochrome P450 (CYP450) enzymes in rat' s heart. <b>Method::Male Sprague Dawley rats were randomly divided into the control group, the inducer group (0.08 g·kg^-1·d^-1), the aconite group (0.5 g·kg^-1·d^-1), the licorice group (0.5 g·kg^-1·d^-1) and the compatibility group (aconite and licorice group 0.5 g·kg^-1·d^-1), with 6 rats in each group. In each group, the serum concentrations of aspartate transaminase (AST), creatine kinase (CK) andlactate dehydrogenase (LDH) were respectively measured. Hematoxylin-eosin(HE) staining was used to detect the pathologic changes in the heart tissue of the rats. The expression of several CYP genes were measured. <b>Result::After combination, the contents of AST, CK and LDH were lower than those of the aconite group. At the same time, the pathological results showed cardiac injury in rats in the aconite group, and licorice could alleviate the cardiac injury caused by aconite. At the level of gene transcription, the glycyrrhizae group could up-regulate the expressions of CYP2C11 and CYP2J3 to different degrees (<italic>P</italic><0.05), and the combination of licorice and aconite could up-regulate the expression of CYP2B1, CYP2C11 and CYP2J3 family (<italic>P</italic><0.05). It suggested that the combination could promote the formation of EETs in arachidonic acid(AA) metabolism, and the aconite could up-regulate the expressions of CYP4A1, CYP4A3, CYP4F5 and CYP4F6 in CYP4 family (<italic>P</italic><0.05), promote the formation of 20-hydroxyeicosatetraenoic acid(20-HETE). The combination of aconite and licorice can weaken the ability of aconite, down-regulate the expression of CYP4A3, CYP4F1, CYP4F5 and CYP4F6 family mRNA (<italic>P</italic><0.05), inhibit the formation of 20-HETE and reduce the cardiotoxicity caused by aconite. <b>Conclusion::The combination of licorice and aconite can regulate the expression of CYP450 enzyme in the heart, increase the production of EETs and the content of 20-HETE, and finally play a role in reducing toxicity.

Effect of Pudilan Xiaoyan Oral Liquid on Acute Lung Injury Rat Induced by Lipopolysaccharide / 中国实验方剂学杂志

Objective:To explore the effect and mechanism of Pudilan Xiaoyan oral liquid(PDL) on the acute lung injury rat induced by lipopolysaccharide (LPS). Method:The 72 Wistar rats were randomly divided into control group, model group, dexamethasone group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group according to body weight.The acute lung injury model was made through inhalation with lipopolysaccharide in the model group, hexadecadrol group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group.To examining each rat alveolar lavage fluid (BALF) of the total number of white blood cells, enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of nuclear transcription factors-kappa B (NF-κB) and interleukin-10 (IL-10).Hematoxylin-eosin(HE) staining was used to observe morphological changes of lung tissue and explore different doses of PDL effect on acute lung injury in rats. Result:Compared with model group, the account of leukocyte in BALF decreased significantly in PDL 7 g·kg-1·d-1 group and PDL 3.5 g·kg-1·d-1 group (PκB significantly decreased in PDL 7, 3.5, 1.75 g·kg-1·d-1 group (P-1·d-1 group (P-1·d-1 group, the inflammation, edema and congestion in lung tissue reduced (PConclusion:PDL has a significant protective effect on the inflammation of acute lung injury model, and its mechanism is related to the expressions of NF-κB and IL-10. PDL could also repair the injury of lung in acute lung injury model.

Pharmacodynamic Evaluation of Tanreqing Inhalation Solution / 中国实验方剂学杂志

Objective: To clarify the antitussive, expectorant, antipyretic and anti-inflammatory effects of Tanreqing inhalation solution, and provide basis and data support for further research and development of this preparation. Method: The methods of cough induced by ammonia and tracheal phenol red excretion were used to observe the antitussive and expectorant effects of Tanreqing inhalation solution in mice. The fever model of rats was established by intraperitoneal injection of bacterial lipopolysaccharide(LPS) to observe the antipyretic effect of the Tanreqing inhalation solution, the acute pneumonia model of rats was established by atomizing LPS inhalation, and the anti-inflammatory effect of Tanreqing inhalation solution was observed. Result: Tanreqing inhalation solution could reduce the number of coughs in mice induced by ammonia water, increase the amount of phenol red excretion in mouse trachea, decrease the levels of body temperature and its related regulatory factors of prostaglandin E2(PGE2) and cyclic adenosine monophosphate(cAMP) of rats induced by LPS, decrease the white blood cell(WBC) count and the neutrophil ratio(NEUT) in bronchoalveolar lavage fluid(BALF) of rats with LPS-induced acute pneumonia, and reduce the levels of nuclear transcription factor-κB(NF-κB) and interleukin-1β(IL-1β) in lung tissue. Conclusion: Tanreqing inhalation solution has obvious antitussive, expectorant, antipyretic and anti-inflammatory effects, which is worthy of further development and promotion.

Effect of Different Administration Methods with Bleomycin on Pulmonary Fibrosis in Rats / 中国实验方剂学杂志

Objective:Compare the effects of 3 administration methods (tracheal perfusion, tail vein injection and aerosol inhalation) with bleomycin (BLM) in inducing pulmonary fibrosis in rats, in order to find out the optimal administration methods. Method:Eighty sprague-dawley (SD) male rats with SPF were randomly divided into aerosol inhalation blank group, single tracheal perfusion group(10 mg·kg-1), multiple tracheal perfusion group(5 mg·kg-1), single intravenous injection group(150 mg·kg-1), multiple intravenous injection group(50 mg·kg-1), single aerosol inhalation group (30 min)and multiple aerosol inhalation group(30 min). The mortality and body weight of rats in each group were observed at 7 d, 14 d and 28 d after the administration. And 28 days later after the administration, the lung coefficients of rats in each group were observed, paraffin sections were prepared, hematoxylin-eosin staining (HE) and Masson staining were performed, and the contents of hydroxyproline (HYP) and plasminogen activator inhibitor-1 (PAI-1) in lung tissues were detected by enzyme-linked immunosorbent assay (ELISA), so as to evaluate the alveoli inflammation and pulmonary fibrosis of rats in each group. Result:Compared with the aerosol inhalation blank group, the rats in the trachea perfusion group had the highest mortality among the drug treatment groups. The pulmonary coefficients of rats in the multiple intravenous injection group and the multiple inhalation group were significantly higher than those in the blank group(PPPConclusion:Bleomycin was inhaled repeatedly to establish pulmonary fibrosis model. The pathological injury and physiological indexes of the model rats were relatively stable, which conforms with the evolution process of pulmonary fibrosis.

Investigation of Physicochemical Properties and Determination Methods of Liposomal Entrapment Efficiency of Dichroa Alkali Hydrochloride / 中国实验方剂学杂志

Objective: To study on the physical and chemical properties of dichroa alkali hydrochloride and to establish a method for the determination of entrapment efficiency of dichroa alkali hydrochloride liposomes. Method: HPLC was used to determine the content of dichroa alkali hydrochloride with mobile phase of acetonitrile-water-triethylamine-glacial acetic acid(9:91:0.35:0.75) and detection wavelength at 265 nm.The oil-water partition coefficient of this compound in different pH range was measured by shake flask method.The stability of the dichroa alkali hydrochloride in phosphate buffer solution with different pH after sterilization at 125℃ for 30 min was investigated.Ammonium sulfate gradient method was used to prepare dichroa alkali hydrochloride liposomes.The microcolumn was prepared by dextran gel and cation exchange resin,respectively.Then the free drug and liposome were separated by centrifugation,the drug content was measured,and the encapsulation efficiency was calculated.The t-test was performed using SPSS 20.0 software,the differences between these two methods were compared. Result: In the pH 6-9,the oil-water partition coefficient of dichroa alkali hydrochloride increased with increasing of pH,which was between 0.016 and 1.44;the recovery rate of dichroa alkali hydrochloride after sterilization was 37.16%-57.91%.Between the dextran gel microcolumn centrifugation and the cation exchange resin microcolumn centrifugation,there was no significant difference in the entrapment efficiency of the liposomes. Conclusion: Dichroa alkali hydrochloride is suitable for preparation of liposomes.However,its stability is not ideal,so the experimental temperature should be strictly controlled in the preparation process.Dextran gel microcolumn centrifugation and cation exchange resin microcolumn centrifugation can be used to determine the entrapment efficiency of dichroa alkali hydrochloride liposomes,and the cation exchange resin microcolumn centrifugation is suggested after comparison.

Protective Effect of Renshen Sinitang and Its Active Ingredients on Myocardial Cell Injury Induced by Pentobarbital Sodium / 中国实验方剂学杂志

Objective: To explore the protective effect and mechanisms of Renshen Sinitang and its active ingredients on cardiomyocyte injury induced by pentobarbital sodium. Method: H9C2 cells were sub-cultured with ginsenoside Rb2 0.01, 0.1, 1 μmol ·L-1, Re 0.01, 0.1, 1 μmol·L-1, isoliquiritigenin 20, 40, 80 μmol·L-1, glycyrrhetinic acid 10, 20, 40 μmol·L-1, Renshen Sinitang, 10, 100, 400 mg·L-1, for 4 h. After treatment with 0.1% of sodium pentobarbital for 30 min, cell viability, lactate dehydrogenase (LDH), lipid peroxide malondialdehyde (MDA), Na+-K+-adenosine triphosphate(ATP) ase, Ca2+-ATPase activity, and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect the expressions of peroxisome proliferative activated receptor-1α (PGC-1α), B-cell lymphoma-2 associated X protein(Bax) and cysteine aspartate-specific protease-3(Caspase-3) mRNA. Result: Renshen Sinitang and its active ingredients have a protective effect on heart failure cell model. Compared with the normal group, the cell survival rate of the model group decreased significantly, while the LDH and MDA contents increased significantly, and the Na+-K+-ATPase activity increased. Ca2+-ATPase activity was significantly decreased, PGC-1α mRNA expression was down-regulated, Bax and Caspase-3 mRNA expressions indicates the modeling(P+-K+-ATPase activity, increased Ca2+-ATPase activity, up-regulated PGC-1α mRNA expression, and inhibited Bax and Caspase-3 mRNA expression (PPConclusion: Renshen Sinitang and its active ingredients have a significant protective effect on heart failure cell model, and its mechanisms of action are related to anti-oxidation, improvement of mitochondrial energy metabolism and inhibition of mitochondrial apoptosis pathway.

Construction and verification of anti-MM scFv-tP fusion protein expression vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct an expression vector of anti-MM scFv-tP fusion protein and test its expression efficiency and function.</p><p><b>METHODS</b>The truncated protamine (tP) gene sequence was added to the gene of single chain antibody against the specific antigen on the surface of malignant melanoma tumor cells using PCR. A GST-fusion expression vector was constructed and the soluable protein was expressed in the E.coli system. After cleavage and purification, the purified fusion protein was obtained. The binding activity of Anti-MM scFv-tP and siRNA was detected by EMSA. Flow cytometry and confocal microscopy were used to detect the cell surface antigen binding activity of the fusion protein.</p><p><b>RESULTS</b>The expression vector of Anti-MM scFv-tP fusion protein was successfully constructed. The soluable protein could be expressed in the E.coli system, and the purified fusion protein was obtained. The anti-MM scFv-tP fusion protein retained siRNA binding ability and could directly target malignant melanoma (MM) LiBr cells.</p><p><b>CONCLUSION</b>The recombinant GST- Anti-MM-scFv-tp expression vector was successfully constructed. The fusion protein retains siRNA binding ability and can directly target LiBr cells to provide a reliable tool for further study.</p>

Dissolving mechanism of strain P17 on insoluble phosphorus of yellow-brown soil

Strain P17 was a bacterial strain identified as Bacillus megaterium isolated from ground accumulating phosphate rock powder. The fermentation broth of strain P17 and the yellow-brown soil from Nanjing Agricultural University garden were collected to conduct this study. The simulation of fixed insoluble phosphorous forms after applying calcium superphosphate into yellow-brown soil was performed in pots, while available P and total P of soil were extremely positive correlative with those of groundwater. Then the dissolving effect of strain P17 on insoluble P of yellow-brown soil was studied. Results showed that Bacillus megaterium strain P17 had notable solubilizing effect on insoluble phosphates formed when too much water-soluble phosphorous fertilizer used. During 100 days after inoculation, strain P17 was dominant. Until the 120th day, compared with water addition, available P of strain P17 inoculation treated soil increased by 3 times with calcium superphosphate addition. Besides available P, pH, activity of acid and alkaline phosphatase and population of P-solubilizing microbes were detected respectively. P-solubilizing mechanism of P-solubilizing bacteria strain P17 seems to be a synergetic effect of pH decrease, organic acids, phosphatase, etc.