Design, Synthesis, Antibacterial, and Antifungal Evaluation of Phenylthiazole Derivatives Containing a 1,3,4-Thiadiazole Thione Moiety.

To effectively control the infection of plant pathogens, we designed and synthesized a series of phenylthiazole derivatives containing a 1,3,4-thiadiazole thione moiety and screened for their antibacterial potencies against Ralstonia solanacearum, Xanthomonas oryzae pv. oryzae, as well as their antifungal potencies against Sclerotinia sclerotiorum, Rhizoctonia solani, Magnaporthe oryzae and Colletotrichum gloeosporioides. The chemical structures of the target compounds were characterized by 1H NMR, 13C NMR and HRMS. The bioassay results revealed that all the tested compounds exhibited moderate-to-excellent antibacterial and antifungal activities against six plant pathogens. Especially, compound 5k possessed the most remarkable antibacterial activity against R. solanacearum (EC50 = 2.23 µg/mL), which was significantly superior to that of compound E1 (EC50 = 69.87 µg/mL) and the commercial agent Thiodiazole copper (EC50 = 52.01 µg/mL). Meanwhile, compound 5b displayed the most excellent antifungal activity against S. sclerotiorum (EC50 = 0.51 µg/mL), which was equivalent to that of the commercial fungicide Carbendazim (EC50 = 0.57 µg/mL). The preliminary structure-activity relationship (SAR) results suggested that introducing an electron-withdrawing group at the meta-position and ortho-position of the benzene ring could endow the final structure with remarkable antibacterial and antifungal activity, respectively. The current results indicated that these compounds were capable of serving as promising lead compounds.

Design, Synthesis, and Antifungal Activity of Some Novel Phenylthiazole Derivatives Containing an Acylhydrazone Moiety.

Crop fungal diseases pose a serious threat to global crop production and quality. Developing new and efficient fungicides is an important measure to control crop diseases. Phenylthiazole was found to be an excellent antifungal skeleton based on our previous study on the structural optimization and biological activity of the natural product thiasporine A. To find new fungicides, 45 phenylthiazole derivatives containing an acylhydrazone moiety were designed and synthesized by the principle of active substructure splicing. Forty-two of the forty-five compounds are novel, except for compounds E1, E14, and E33. Their structures were structurally characterized by 1H NMR, 13C NMR, and HRMS. The antifungal activities of the target compounds against Magnaporthe oryzae Colletotrichum camelliaet, Bipolaris maydis, and Sclerotinia sclerotiorum were evaluated at 25 µg/mL. The bioassay results revealed that most of these compounds exhibited excellent antifungal activities against M. oryzae and C. camelliaet at 25 µg/mL. In particular, compounds E4, E10, E14, E17, E23, E26, and E27 showed the inhibition rate of more than 80% against M. oryzae, with EC50 values of 1.66, 2.01, 2.26, 1.45, 1.50, 1.29, and 2.65 µg/mL, respectively, which were superior to that of the commercial fungicides Isoprothiolane (EC50 = 3.22 µg/mL) and Phenazine-1-carboxylic acid (EC50 = 27.87 µg/mL). The preliminary structure-activity relationship (SAR) results suggested that introducing methyl, halogen, or methoxy at the ortho-position of R1 and the para-position of R2 can endow the final structure with excellent antifungal activity against M. oryzae. The current results provide useful data for developing phenylthiazole derivatives as new fungicides for controlling rice blast caused by M. oryzae.

Design, synthesis and antifungal evaluation of phenylthiazole-1,3,4-oxadiazole thione (ketone) derivatives inspired by natural thiasporine A.

BACKGROUND: Plant pathogenic fungal infections have become a severe threat to the yield and quality of agricultural products, and new green antifungal agents with high efficiency and low toxicity are needed. In this study, a series of thiasporine A derivatives containing phenylthiazole-1,3,4-oxadiazole thione (ketone) structures were designed and synthesized, and their antifungal activities against six invasive and highly destructive phytopathogenic fungi were evaluated. RESULTS: The results found that all compounds showed moderate to potent antifungal activity against six phytopathogenic fungi, and most of the E series compounds showed remarkable antifungal activity against Sclerotinia sclerotiorum and Colletotrichum camelliaet. In particular, compounds E1-E5, E7, E8, E13, E14, E17, and E22 showed more significant antifungal activity against S. sclerotiorum, with half-maximal effective concentration (EC50 ) values of 0.22, 0.48, 0.56, 0.65, 0.51, 0.39, 0.60, 0.56, 0.60, 0.63, and 0.45 µg mL-1 , respectively, which were superior to that of carbendazim (0.70 µg mL-1 ). Further activity studies showed that compound E1 possessed superior curative activities against S. sclerotiorum in vivo and better inhibitory effects on sclerotia germination and the formation of S. sclerotiorum compared with those of carbendazim. CONCLUSIONS: This study indicates that these thiasporine A derivatives containing phenylthiazole-1,3,4-oxadiazole thione structures might be used as antifungal agents against S. sclerotiorum. © 2023 Society of Chemical Industry.

Rhein-Amino Acid Ester Conjugates as Potential Antifungal Agents: Synthesis and Biological Evaluation.

In the search for crop protectants, amino acid ester conjugates have been widely investigated as potential antifungal agents. In this study, a series of rhein-amino acid ester conjugates were designed and synthesized in good yields, and their structures were confirmed by 1H-NMR, 13C-NMR and HRMS. The bioassay results revealed that most of the conjugates exhibited potent inhibitory activity against R. solani and S. sclerotiorum. In particular, conjugate 3c had the highest antifungal activity against R. solani with an EC50 value of 0.125 mM. For S. sclerotiorum, conjugate 3m showed the highest antifungal activity with an EC50 value of 0.114 mM. Satisfactorily, conjugate 3c exhibited better protective effects than that of the positive control, physcion, against powdery mildew in wheat. This research supports the role of rhein-amino acid ester conjugates as potential antifungal agents for plant fungal diseases.

Effect of personalized nursing based on quantitative evaluation strategies on remission of first-episode schizophrenia comparing to usual care: a systematic review and meta-analysis.

BACKGROUND: The drug control of symptoms is for now the main clinical treatment of schizophrenia, but patients' varying condition and poor compliance can also fluctuate the therapeutic effect. Personalized nursing with a quantitative evaluation strategy (PNQES) may help improve the compliance and symptoms, but there are controversies over the outcomes reported in each specific study; the meta-analysis method aims to resolve the controversies over studies, thus, we conducted this study to pooling the results of controlled clinical studies, and to systematically evaluate the effects of this nursing model. METHODS: The PubMed, Medline, Embase, China National Knowledge Infrastructure, and Wanfang databases were selected and searched for relevant articles for PNQES comparing to usual care. The inclusion criteria were established according to the Participants, Interventions, Comparisons, Outcomes, and Study (PICOS) framework. The Cochrane risk of bias 2.0 tool was used to evaluate the risk of bias of the included articles. The symptom scores, treatment compliance rate, quality of life, and social function indicators of the patients after nursing were quantitatively analyzed with effect sizes of mean difference (MD) or standard mean difference (SMD). RESULTS: The 11 included articles comprised a total of 1,251 patients with experimental group 625 and control group 626. Of all the 11 articles, only 1 had a "low" risk of bias, while the other articles had "some concern of risk;" none of the articles had a "high" risk of bias. The meta-analysis showed that patients who received PNQES had a significantly lower Positive and Negative Syndrome Scale (PANSS) total score after care than patients who received routine care [MD =-9.95, 95% confidence interval (CI): -14.35, -5.55; P<0.00001]. Further, the treatment compliance rate of patients who received PNQES was significantly higher (odds ratio =4.44, 95% CI: 2.17, 9.09; P<0.0001), as was the quality of life (standard MD =2.40, 95% CI: 1.46, 3.34; P<0.00001). Further, the social function deficit score was significantly lower (MD =-2.25, 95% CI: -3.75, -0.76; P=0.003). Subgroup and regression analyses showed that patient age, initial PANSS score, and the quantitative method of disease severity were not the sources of heterogeneity. Different intervention approaches applied may have been the source of heterogeneity. DISCUSSION: The application of PNQES is helpful for improving patients' symptoms and disease outcomes, treatment compliance, social function, and quality of life. It is suggested to be generalized in clinical application.

The Effect of Social Cognitive Interaction Training on Schizophrenia: A Systematic Review and Meta-Analysis of Comparison with Conventional Treatment.

Background: Existing antipsychotic medications may alleviate the majority of patients' symptoms, but they have no discernible impact on improving social function and quality of life. Psychotherapy is required for the treatment of schizophrenia. However, contemporary psychotherapy technology intervention techniques are limited to a single intervention, and there is a lack of holistic and complete intervention approaches. Social cognition and interaction training is a comprehensive therapy strategy that has been employed in clinical practice; however, the therapeutic efficacy has been inconsistently reported. As a result, we included controlled clinical trials for meta-analysis in order to carefully assess the efficacy of this therapy. Methods: This meta-analysis searched all RCT literatures related to social cognitive interaction training (SCIT) published before April 2022 and assessed the effect of this method in the treatment of schizophrenia. The data in the literatures were combined, and the standardized mean difference (SMD) and mean difference (MD) with 95% confidence interval (95% CI) were calculated to predict the negative symptom score, positive symptom score, PANSS score, and social function score of the patients after treatment. Results: 14 RCT studies including 1167 inpatients with schizophrenia were included in this study using a retrospective observational study method, including 590 patients treated with SCIT and 577 patients treated with treatment as usual (TAU). The pooled analysis showed that patients after SCIT had lower negative symptom scores (SMD = -1.66, 95% CI (-2.32, -1.00), P < 0.0001), lower positive symptom scores (MD = -4.03, 95% CI (-7.69, -0.36), P = 0.03), lower PANSS total scores (MD = -6.33, 95% CI (-12.43, -0.23), P = 0.02), and higher social functioning scores (SMD = 0.77, 95% CI (0.34, 1.20), P < 0.001) than those after TAU. Conclusion: Our findings support that SCIT is helpful to improve the relief of symptoms and the improvement of social function in patients with schizophrenia, providing a basis for the application of SCIT in hospitalized patients and community patients, and can guide the treatment and intervention of patients with schizophrenia.

Study on the selectivity and phloem mobility of Fenoxaprop-P amino acid ester conjugates on rice and barnyard grass.

To improve the selectivity of the fenoxaprop herbicide to rice and barnyard grass, a series of fenoxaprop-P-ethyl-amino acid ester conjugates were designed and synthesized, and tested for biological activity as well as their phloem mobility. The bioassay results indicated that the target compounds possessed better activity against barnyard grass (Echinochloa crusgalli) than rape (Brassica campestris L.) at the concentration of 0.5 mmol/L. Compounds 3h and 3i, showed more than 70% control efficiency against barnyard grass, while less than 30% for rape. The compounds showed less impact on rice after spray treatment than in the germination test. Compounds 3i, 3j, and 3k showed excellently herbicidal activities against barnyard grass and low phytotoxicity to rice. Compound 3k showed 6.1% phytotoxicity to rice at a spray concentration of 0.25 mmol/L, better than fenoxaprop-P-ethyl (61.6%) at the same concentration. The selectivity results of the target compounds revealed that most of compounds obviously reduced phytotoxicity to rice while retaining herbicidal activity of barnyard grass. The herbicidal activity of compound 3d compared to FPE was increased by 50%, while its safety on rice was also increased by 50%. The concentration of the compounds in barnyard grass roots was higher than in rice roots, showing greater phloem mobility. In particular, the concentration of compound 3d on barnyard grass exhibited 142.72 mg/kg which was 3 times as much as Fenoxaprop, while its concentration on rice exhibited 3.65 mg/kg, the results revealed that the difference of phloem mobility might be the important reason for causing the selectivity.

The influence of steric configuration of phenazine-1-carboxylic acid-amino acid conjugates on fungicidal activity and systemicity.

BACKGROUND: Conjugating an amino acid onto existing fungicidal parent structures has been demonstrated to be an effective way to endow non-phloem mobile fungicides with phloem mobility. To alter the systemicity of the fungicide PCA (phenazine-1-carboxylic acid), 10 amino acids derivatives of this fungicide were designed and synthesized, and their synthesis, characterization, phloem and xylem mobility in Ricinus communis L, and their fungicidal activity in vitro are described. RESULTS: The systemicity experiments in Ricinus communis system demonstrated that all conjugates exhibited obvious phloem mobility compared with non-phloem-mobile PCA, and the introduction of an L-amino acid to PCA more greatly enhanced the phloem mobility. The five D-amino acid conjugates exhibited higher xylem mobility than that of PCA and of each corresponding L-amino acid conjugate. Most conjugates were found to exhibit moderate in vitro fungicidal activities against six pathogenic fungi, which were lower than that of PCA. The results of the bioassay showed fungicidal activities of PCA-amino acid conjugates associated not only with different amino acids, but also with their conformation. Conjugation with D-amino acid contributed to the in vitro fungicidal activities of PCA-amino acid conjugates. CONCLUSIONS: The current research offers a new strategy for enhancing the systemicity of non-phloem-mobile fungicides and presents some useful information on the effects of introducing amino acids of different steric configurations on the fungicidal activity, phloem and xylem mobility of the parent fungicide. © 2019 Society of Chemical Industry.

Synthesis and bioactivities of diamide derivatives containing a phenazine-1-carboxamide scaffold.

Taking natural product phenazine-1-carboxamide (PCN) as a lead compound, a series of novel phenazine-1-carboxylic acid diamide derivatives were designed and synthesised. Their structures were confirmed by 1H-NMR and HRMS. The bioassays showed that some of the target compounds exhibited promising in vitro fungicidal activities, and exhibited excellent and selective herbicidal activities. Particularly, compounds c, h, o and s displayed root length inhibition activities against barnyard grass with the rate of more than 80%. Compound c exhibited the best activity among all the target compounds against barnyard grass stalk length with the IC50 value of 0.158 mmol/L, and compound o exhibited the best and wide spectrum inhibition against barnyard grass root length and rape in both root length and stalk length herbicidal activities with its IC50 values of 0.067, 0.048 and 0.059 mmol/L respectively. The analysis of preliminary Structure-Activity Relationships provides the theoretical basis for further design of phenazine-1-carboxylic acid.

Design, synthesis and biological activity of hydroxybenzoic acid ester conjugates of phenazine-1-carboxylic acid.

We prepared 16 novel hydroxybenzoic acid ester conjugates of phenazine-1-carboxylic acid (PCA) and investigated their biological activity. Most of the synthesized conjugates displayed some level of fungicidal activities in vitro against five phytopathogenic fungi. Nine conjugates 5b, 5c, 5d, 5e, 5h, 5i, 5m, 5n and 5o (EC50 between 3.2 µg/mL and 14.1 µg/mL) were more active than PCA (EC50 18.6 µg/mL) against Rhizoctonia solani. Especially conjugate 5c showed the higher fungicidal activity against Rhizoctonia solani which is 6.5-fold than PCA. And the results of the bioassay indicated that the fungicidal activity of conjugates was associated with their LogP, and the optimal LogP values of the more potent fungicidal activities within these conjugates ranged from 4.42 to 5.08. The systemic acquired resistance induced by PCA-SA ester conjugate 5c against rice sheath blight disease in rice seedlings was evaluated. The results revealed that PCA-SA ester conjugate 5c retained the resistance induction activity of SA against rice sheath blight.

Design, Synthesis, Phloem Mobility, and Bioactivities of a Series of Phenazine-1-Carboxylic Acid-Amino Acid Conjugates.

Developing fungicides with phloem mobility that can be applied to leaves to control root or vascular pathogens has long been desirable. To achieve this goal, an efficient and economical strategy involves introducing an amino acid into the existing highly active parent pesticide molecule. Hence, 12 L-phenazine-1-carboxylic acid (PCA)-amino acid conjugates 4a⁻l were designed and synthesized via a simple synthetic route. In vitro bioassays results showed that all synthesized compounds 4a⁻l exhibited certain fungicidal activities against six tested fungi. Compound 4c exhibited relatively good fungicidal activity against Rhizoctonia solani, and the EC50 value was 0.084 ± 0.006 mmol/L. The phloem mobility experiments revealed that introducing an amino acid to PCA could effectively endow PCA with phloem mobility in R. communis L. Among them, nine conjugates were found in phloem sap, and L-PCA-Valine 4d exhibited the highest phloem mobility. Analysis results from the prediction of the Kleier model indicated that an active carrier-mediated mechanism may be involved in L-PCA-amino acid conjugates-a result that needs to be confirmed and complemented with further tests. The current research provides useful data for modifying non-phloem-mobile fungicidal molecules to phloem-mobile types.

Synthesis, fungicidal activity and phloem mobility of phenazine-1-carboxylic acid-alanine conjugates.

Phenazine-1-carboxylic acid (PCA) is a natural product that has been proven effective against a number of soil-borne fungal phytopathogens and registered for biofungicide against rice sheath blight in China. In order to improve the phloem mobility of phenazine-1-carboxylic acid (PCA), four PCA derivatives were designed and synthesized by conjugating PCA with l-alanine methyl ester, d-alanine methyl ester, l-alanine and d-alanine respectively. In vitro and planta bioassays results showed that conjugates L-PAM and D-PAM exhibited higher fungicidal activities against Rhizoctonia solani Kuhn than PCA while L-PA and D-PA were less active than PCA. The concentration of conjugates in Ricinus communis phloem sap was determined by HPLC. The results showed that only L-PA exhibited phloem mobility among these conjugates, and its concentration in Ricinus communis phloem sap increased with the increase of time (the maximum concentration was 12.69µM within 5h). However, the results of pot experiments showed that L-PA and other conjugates didn't exhibited the inhibition for the growth of Rhizoctonia solani Kuhn in the lower leaves after treatment in the upper leaves of rice seedlings. This may be due to the poor plant absorbility for them or their too little amount of accumulation in the lower leaves.

Podocyte injury and overexpression of vascular endothelial growth factor and transforming growth factor-beta 1 in adriamycin-induced nephropathy in rats.

The aim of this study is to investigate the expression of nephrin, vascular endothelial growth factor (VEGF), transforming growth factor-beta 1 (TGF-ß1), and podocyte number in adriamycin (ADR)-induced nephropathy. A total of 60 male Sprague-Dawley rats were randomly divided into the control group and the ADR nephropathy group. The nephropathy was induced by tail-vein injection of ADR (4 mg/kg) twice at a 14-day interval. The expression levels of nephrin, VEGF, and TGF-ß1 in glomeruli were assessed by immunohistochemistry and western blotting. The podocyte number was also evaluated after anti-Wilms' tumor-1 (WT1) immunohistochemical staining. In addition, the urinary protein content, biochemical parameters in serum samples and glomerular sclerosis index (SI) were compared between groups. In the ADR nephropathy group, the expression levels of nephrin was significantly decreased with the fusion of podocyte foot processes at 6 weeks after the first ADR injection, which was associated with a marked proteinuria. A decrease in podocyte number and an increase in SI with the overexpression of both VEGF and TGF-ß1 were also observed in the glomeruli at 10 weeks after the first ADR injection. This was associated with focal segmental glomerulosclerosis (FSGS). The study data suggest that podocyte injury and decreased nephrin, as well as increased VEGF and TGF-ß1, may contribute to the development of proteinuria and FSGS in ADR-induced nephropathy in rats.

[Diabetes mellitus ulcers treatment with Bletilla striata polysaccharide].

The aim of this study was to evaluate the efficacy of Bletilla striata polysaccharide on diabetes mellitus ulcers. Diabetes mellitus animal model was established by single ip injection of streptozotocin (STZ, 50 mg x kg(-1)) with the criteria of blood glucose > or = 16.7 mmol x L(-1) after 72 h. 4 weeks after STZ injection, each animal received two full thickness incisional wounds (1.8 cm in diameter). The wounds then were divided into B. striata polysaccharide group and PBS group. Wound closure rate, fibroblast (FB) infiltration, hydroxyproline (OHP) content and myeloperoxidase (MPO) levels were examined on day 3, 7, 14, 21 post wound. The treatment of B. striata polysaccharide significantly facilitated diabetes mellitus ulcers healing compared to PBS group. Histological analysis showed that B. striata polysaccharide markedly increased inflammatory cell infiltration in wound area. The herb also strongly evaluation of FB, OHP demonstrated a significantly increased in B. striata polysaccharide group. B. striata polysaccharide group promoted wound closure by means of enhanced inflammatory cell infiltration and re-epithelialization, and the promotion of FB and OHP levels.

[Study on effect of total saponins from Semen Nigellae on inflammatory mediators and ERK/MAPK pathway in stimulated macrophage].

OBJECTIVE: To study the anti-inflammatory mechanism of total saponins from Semen Nigellae (TSSN). METHOD: IFN-gamma plus LPS stimulated RAW 264. 7 macrophage has been used as inflammatory experimental model. Griess reaction for nitric oxide production, FRAP assay for total antioxidant capacity, RT-PCR for mRNA expression and Western blot for protein expression examination were performed. RESULT: TSSN inhibited NO production in a dose-dependent manner. The gene and protein expression of iNOS were also suppressed by the herb extract. TSSN treatment significantly attenuated mRNA of inflammatory mediators such as COX-2, IL-1beta, IL-6 while increased PPAR-gamma gene and protein expression. Furthermore, phosphorylation of ERK (p-ERK) was markedly inhibited by TSSN. CONCLUSION: TSSN suppressed pro-inflammatory mediators such as COX-2, IL-1beta, IL-6 and increased anti-inflammatory mediator PPAR-gamma expression. Meanwhile, TSSN inhibited over production of NO and iNOS expression through ERK/MAPK pathway.

Developing a system for regulating expression of human hepatocyte growth factor using tetracycline in NRK52E cells.

INTRODUCTION: Hepatocyte growth factor (HGF) is a target of gene therapy for renal fibrosis. The aim of this study was to establish a human HGF gene expression system that is regulated by tetracycline (Tet) in normal rat kidney tubular epithelial cells (NRK52E cells). MATERIALS AND METHODS: The plasmids pTet-on, pBI-L-HGF and pTK-Hyg were transfected sequentially into NRK52E cells using Lipofectamine 2000. The expression of HGF gene was measured, and the activity of expressed HGF was detected. RESULTS: A clone of pBI-L-HGF/NRK52E cells showing strong reaction to doxycycline (Dox) was selected using a luciferase reporter assay system. The expression of both HGF mRNA and protein was significantly higher (both p < 0.01) in the Dox group than that in the control group. Furthermore, the bioactivity of expressed HGF was confirmed in the assay. CONCLUSIONS: A Tet-regulated human HGF gene expression system in NRK52E cells has been established. This cell line may prove useful for gene therapy against renal fibrosis.

[Podocyte injury and expression of nephrin and VEGF in rat nephrosis model induced by adriamycin].

OBJECTIVE: To investigate podocyte injury and the expression of nephrin and VEGF in rat nephrosis model induced by adriamycin. METHODS: The rat adriamycin induced nephrosis model was established, while the biochemical indicators in blood and urine were measured and the pathological changes of the renal tissue were evaluated by light microscope and electron microscope. The podocyte number was counted, and the expression levels of nephrin, VEGF were examined at different time by means of immunohistochemistry. RESULTS: After second injected with adriamycin,the model group nephrin presented a weak signal in the end of the first week (P < 0.05), and the expression of VEGF started to increase at the end of the eighth week (P < 0.05). The podocyte number decreased at the end of the eighth week (P < 0.05). The expression of nephrin and the number of podocyte were negatively correlated with the 24-hour urine protein, blood urea nitrogen and serum creatinine; while the expression of VEGF was positively correlated with the 24-hour urine protein, blood urea nitrogen and serum creatinine. CONCLUSION: The decrease of nephrin expression and the change of its distribution might be the significant factors resulting in considerable proteinuria. VEGF participated in the process of proteinuria and glomerular sclerosis in the development of rat adriamycin nephrosis.

[Construction and identification of tetracycline-inducible human hepatocyte growth factor eukaryotic expression vector].

OBJECTIVE: To construct a tetracycline-inducible eukaryotic expression vector containing human hepatocyte growth factor (HGF) cDNA. METHODS: Human HGF cDNA fragment was obtained by PCR from pUC-SRalpha/HGF plasmid and inserted into the restriction site between Mlu I and Sal I of the tetracycline-inducible eukaryotic expression vector pBI-L. pBI-L-HGF was constructed by DNA recombination in vitro, and was identified by restriction endonucleases digestion and sequencing. RESULTS: The fragment of pBI-L-HGF digested with restriction endonucleases well corresponded to expectation, and the sequence of inserted HGF cDNA was correct according to the GenBank. CONCLUSION: The tetracycline-inducible eukaryotic expression vector of human HGF pBI-L-HGF has been constructed successfully, which allows further study of HGF gene therapy with much safety and easy manipulation.

[Construction and identification of tetracycline-inducible rat Smad7 eukaryotic expression vector].

OBJECTIVE: To construct a tetracycline-inducible eukaryotic expression vector of rat Smad7. METHODS: The total RNA was extracted from normal rat kidney with Trizol agent. Rat Smad7 cDNA fragment was cloned by RT-PCR, and was inserted into the restriction site between Nhe I and Hind III of the inducible eukaryotic expression vector pBI-L by tetracycline. pBI-L-Smad7 was constructed by digestion and ligation, and detected by restriction endonuclease digestion and sequencing. RESULTS: The recombinant eukaryotic expression vector pBI-L-Smad7 was constructed correctly as confirmed by restriction endonuclease digestion and sequencing. The fragment of pBI-L-Smad7 digested with restriction endonucleases and the sequence of inserted Smad7 cDNA were consistent with the results of theoretical analysis. CONCLUSION: The tetracycline- inducible eukaryotic expression vector of rat Smad7, pBI-L-Smad7, is constructed successfully, which may facilitate further clinical study of Smad7 gene therapy for tissue and organ fibrosis.

[The protective effects of three components isolated from Astragalus membranaceus on shock wave lithotripsy induced kidney injury in rabbit model].

OBJECTIVE: Extracorporeal shock wave lithotripsy (ESWL) is has been shown to reduce renal parenchymal injury subject to application of shock wave lithotripsy in our pervious study. To investigate the protective action of three main components from Astragalus membranaceus, including total saponins of astragalus (TSA), total flavonoids of astragalus (TFA) total polysaccharide of astragalus (TPA) in alleviating shock wave induced kidney damage. METHODS: Sixty four male rabbits were randomly assigned to a control group or to 3 groups that were premedicated with TSA TFA and TPA respectively prior to application of ESWL. Each group of animals underwent shock wave lithotripsy (18 kV) to the right kidneys and received a total of 1500 shocks. Peripheral blood samples were collected to evaluate the levels of plasma endothelin-1 (ET-1), plasma nitric oxide (NO) and serum malondialdehyde (MDA) before and after shock wave treatment. The concentrations of these markers in the treated kidney tissues were also detected 3 days, 7 days and 14 days after application of ESWL. The changes of histopathology and cells ultrastructure were observed through light microscope and electron microscope. Untreated contralateral kidneys were evaluated as controls. RESULTS: In control serials the levels of ET-1 and MDA were elevated significantly while the level of NO was significantly decreased after application of shock wave lithotripsy (P < 0.05). The comparison between the controls and premedicated groups demonstrated that all these three components especially TSA and TFA significantly inhibited shock wave induced increasing of ET-1 and MDA (P < 0.05). TSA also significantly suppressed the decrease of NO and made the recovery time earlier compare to the results of controls (P < 0.05). However, TFA and TPA had almost no effects on the change of NO. (P > 0.05). The results in histopathology showed noticeably damage of glomerular and tubular epithelial cells in the treated kidneys in the controls. The histological alterations in the TPA group were similar to those of the controls. These alterations were significantly milder in the TSA and TFA particular the TSA group. CONCLUSION: TFA and TSA, especially TSA seemed to play the key role in alleviating ESWL induced kidney damage.