Metagenomic analysis reveals gene taxonomic and functional diversity response to microplastics and cadmium in an agricultural soil.

Both microplastics (MPs) and heavy metals are common soil pollutants and can interact to generate combined toxicity to soil ecosystems, but their impact on soil microbial communities (e.g., archaea and viruses) remains poorly studied. Here, metagenomic analysis was used to explore the response of soil microbiome in an agricultural soil exposed to MPs [i.e., polyethylene (PE), polystyrene (PS), and polylactic acid (PLA)] and/or Cd. Results showed that MPs had more profound effects on microbial community composition, diversity, and gene abundances when compared to Cd or their combination. Metagenomic analysis indicated that the gene taxonomic diversity and functional diversity of microbial communities varied with MPs type and dose. MPs affected the relative abundance of major microbial phyla and genera, while their coexistence with Cd influenced dominant fungi and viruses. Nitrogen-transforming and pathogenic genera, which were more sensitive to MPs variations, could serve as the indicative taxa for MPs contamination. High-dose PLA treatments (10%, w/w) not only elevated nitrogen metabolism and pathogenic genes, but also enriched copiotrophic microbes from the Proteobacteria phylum. Overall, MPs and Cd showed minimal interactions on soil microbial communities. This study highlights the microbial shifts due to co-occurring MPs and Cd, providing evidence for understanding their environmental risks.

GDSL Esterase/Lipase GELP1 Involved in the Defense of Apple Leaves against Colletotrichum gloeosporioides Infection.

GDSL esterases/lipases are a subclass of lipolytic enzymes that play critical roles in plant growth and development, stress response, and pathogen defense. However, the GDSL esterase/lipase genes involved in the pathogen response of apple remain to be identified and characterized. Thus, in this study, we aimed to analyze the phenotypic difference between the resistant variety, Fuji, and susceptible variety, Gala, during infection with C. gloeosporioides, screen for anti-disease-associated proteins in Fuji leaves, and elucidate the underlying mechanisms. The results showed that GDSL esterase/lipase protein GELP1 contributed to C. gloeosporioides infection defense in apple. During C. gloeosporioides infection, GELP1 expression was significantly upregulated in Fuji. Fuji leaves exhibited a highly resistant phenotype compared with Gala leaves. The formation of infection hyphae of C. gloeosporioides was inhibited in Fuji. Moreover, recombinant His:GELP1 protein suppressed hyphal formation during infection in vitro. Transient expression in Nicotiana benthamiana showed that GELP1-eGFP localized to the endoplasmic reticulum and chloroplasts. GELP1 overexpression in GL-3 plants increased resistance to C. gloeosporioides. MdWRKY15 expression was upregulated in the transgenic lines. Notably, GELP1 transcript levels were elevated in GL-3 after salicylic acid treatment. These results suggest that GELP1 increases apple resistance to C. gloeosporioides by indirectly regulating salicylic acid biosynthesis.

Trichoderma longibrachiatum T6: A nematocidal activity of endochitinase gene exploration and its function identification.

Endochitinase is a natural extracellular protein in Trichoderma longibrachiatum T6, which can degrade the eggshell of Heterodera avenae significantly, however the related genes that coding this protein was rarely characterized. In the present study, the endochitinase 18-5 gene (T6-Echi18-5) of T. longibrachiatum T6 was cloned and sequenced. The expression level of T6-Echi18-5 gene in T. longibrachiatum T6 was induced and increased after the H. avenae cysts inoculation. The full-length cDNA sequence of T6-Echi18-5 was 1671 bp that contained an ORF of 1275 bp, corresponding to 424 amino acids with a 45.9 kDa molecular weight. A single band of 60.04 kDa was detected and identified using SDS-PAGE and Western blot analysis after transferring the T6-Echi18-5 gene to Escherichia coli BL21 Rosetta (DE3). The concentration of purified recombinant T6-Echi18-5 protein was 1.53 mg·ml-1, and the optimal temperature and pH were 50 °C and 5.0, respectively. The eggshell and content were dissolved and exuded from 4 to10 days after treatment with the purified recombinant T6-Echi18-5 protein. The relative inhibition rate of eggs hatching was 86.79 % at 12 days after treatment. Our study demonstrated the key role of T6-Echi18-5 gene in degrading the H. avenae eggshell and inhibiting the eggs hatching.

Contribution of Nano-Zero-Valent Iron and Arbuscular Mycorrhizal Fungi to Phytoremediation of Heavy Metal-Contaminated Soil.

Soil pollution with heavy metals has attracted increasing concern, which calls for the development of new remediation strategies. The combination of physical, chemical, and biological techniques can achieve more efficient remediation. However, few studies have focused on whether nanomaterials and beneficial microbes can be jointly used to facilitate phytoremediation. Therefore, we studied the role of nano-zero-valent iron (nZVI) and arbuscular mycorrhizal (AM) fungi in the phytoremediation of an acidic soil polluted with Cd, Pb and Zn, using sweet sorghum. X-ray diffraction (XRD), energy dispersive X-ray spectroscopy (EDS), and mapping analyses were conducted to explore the mechanisms of metal immobilization by nZVI. The results showed that although both bare nZVI (B-nZVI) and starch-stabilized nZVI (S-nZVI) inhibited root mycorrhizal colonization, Acaulospora mellea ZZ successfully colonized the plant roots. AM inoculation significantly reduced the concentrations of DTPA-Cd, -Pb, and -Zn in soil, and the concentrations of Cd, Pb, and Zn in plants, indicating that AM fungi substantially facilitated heavy metal immobilization. Both B-nZVI and S-nZVI, ranging from 50 mg/kg to 1000 mg/kg, did not impede plant growth, and generally enhanced the phytoextraction of heavy metals. XRD, EDS and mapping analyses showed that S-nZVI was more susceptible to oxidation than B-nZVI, and thus had more effective immobilization effects on heavy metals. Low concentrations of nZVI (e.g., 100 mg/kg) and AM inoculation had synergistic effects on heavy metal immobilization, reducing the concentrations of Pb and Cd in roots and enhancing root Zn accumulation. In conclusion, our results showed that AM inoculation was effective in immobilizing heavy metals, whereas nZVI had a low phytotoxicity, and they could jointly contribute to the phytoremediation of heavy metal-contaminated soils with sweet sorghum.

Optimization of the Fermentation Media and Parameters for the Bio-control Potential of Trichoderma longibrachiatum T6 Against Nematodes.

The cereal cyst nematode Heterodera avenae is one of the important soil-borne pathogens of cereal crops and causes high yield losses worldwide. Trichoderma spp. formulations are applied as commercial bio-control agents against soil-borne plant pathogens such as H. avenae. However, the relationship between Trichoderma longibrachiatum fermentation parameters and its bio-control potential against H. avenae has not been exclusively established. In the present study, the effect of 10 different fermentation media and conditions on the nematicidal activity of T. longibrachiatum T6 (T6) was evaluated with a single-factor method and a Plackett-Burman design, and the interaction between different fermentation parameters was investigated by a Box-Behnken design. The variables for enhancing the nematicidal activity of T6 culture filtrates were explored and optimized using response surface methodology (RSM). The Minor Medium (MM) plus wheat bran-2 medium was found to be the most effective fermentation medium for T6 culture filtrates against the second stage juveniles (J2s) of H. avenae. The maximum mortality of the J2s was obtained using the T6 culture filtrates under the following fermentation conditions: initial pH 6, 28°C culture temperature, 180 rpm rotating speed, 60 ml of fermentation media, 7 days of incubation time, and 1 ml of inoculation volumes. Among these parameters, the initial pH, inoculation volume, and incubation day were identified as the most significant parameters and critical independent variables for enhancing the nematicidal activity of the T6 culture filtrates. After further optimizations based on statistical predictions, the highest nematicidal activity (92.42%) was obtained with the T6 culture filtrates fermented under an initial pH of 6.06, an inoculation volume of 1.62 ml, and an incubation time of 7.15 days. The nematicidal activity was increased approximately by as high as 1.07% compared with that before optimization. Bio-control efficacy of T6 culture filtrates was 83.88% at the 70th day after wheat seeds sowing in greenhouse experiments. The results from the validation experiments agreed with the model predictions. Our study has improved the bio-control potential of Trichoderma spp. against the plant-parasitic nematodes H. avenae and provided a cost-efficient bio-resource in the future development of novel bio-control agents.

Interactions of microplastics and cadmium on plant growth and arbuscular mycorrhizal fungal communities in an agricultural soil.

Microplastics (MPs) as emerging contaminants have attracted attention worldwide, but little is known on their interactions with metallic contaminants in soil-plant systems. Here, we investigated the interactions between MPs, i.e., polyethylene (PE) and polylactic acid (PLA), and cadmium (Cd) on plant performance and arbuscular mycorrhizal fungal community in an agricultural soil. PE showed no noticeable phytotoxicity, while 10% PLA decreased maize biomass and chlorophyll content in leaves. A significant interaction on root biomass occurred between PE and Cd, but not between PLA and Cd. Both PE and PLA caused increase in soil pH and DTPA-extractable Cd concentrations, but no alterations in Cd accumulation in plant tissues. Different numbers of endemic and total OTUs were observed in various treatments. The relative abundance of arbuscular mycorrhizal fungi (AMF) genera highly varied with MPs and Cd. MPs altered AMF community structure and diversity, depending on their type and dose. Coexisting Cd produced slight but significant interactions with MPs on the dominant AMF genera. Overall, plant growth and AMF community varied with MPs type and dose, Cd, and their interactions, and the high dose of PLA produced stronger phytotoxicity. In conclusion, coexisting MPs and Cd can jointly drive shifts in plant performance and root symbiosis, thereby posing additional risks for agroecosystems and soil biodiversity.

Remediation of Cr(VI)-Contaminated Soil by Nano-Zero-Valent Iron in Combination with Biochar or Humic Acid and the Consequences for Plant Performance.

Nano-scale zero-valent iron (nZVI) is among the most common nanoparticles widely used for the treatment of various environmental contaminants. However, little is known about the combined effects of nano-zero-valent iron (nZVI) and other soil amendments on soil remediation and plant performance. For the first time, we studied the remediation of Cr(VI)-contaminated soil using bare nZVI (B-nZVI) and starch-supported nZVI (S-nZVI) in combination with either biochar (BC) or humic acid (HA), and the consequent effects on plant growth and Cr accumulation. Both S-nZVI and B-nZVI decreased the contents of Cr(VI) and available Cr in soil, but increased available Fe content, with S-nZVI generally showing more pronounced effects at a higher dose (1000 mg/kg). B-nZVI exerted no inhibition and even stimulation on plant growth, but 1000 mg/kg S-nZVI produced significant phytotoxicity, resulting in decreased plant growth, low chlorophyll content in leaves, and excessive accumulation of Fe in roots. Each nZVI decreased shoot and root Cr concentrations. BC and HA produced synergistic effects with nZVI on Cr(VI) removal from soil, but HA decreased soil pH and increased the availability of Cr and Fe, implying a potential environmental risk. Addition of BC or HA did not alter the effects of either nZVI on plant growth. In conclusion, combined application of 100 mg/kg nZVI and BC could be an ideal strategy for the remediation of soil contaminated with Cr(VI), whereas high-dose S-nZVI and HA are not recommended in the remediation of agricultural soils for crop production or in the phytostabilization of Cr(VI).

Phytotoxicity of iron-based materials in mung bean: Seed germination tests.

Environmental applications and potential risks of iron-based materials have attracted increasing attention. However, most previous studies focused on a single material. Comparative research using different iron-based materials under the same experimental conditions is still lacking. Here, six iron-based materials, including micro-sized and nanoscale Fe3O4 (i.e., mFe3O4 and nFe3O4), bulk and bare nanoscale zero-valent iron (i.e., mZVI and B-nZVI), starch-supported nZVI (S-nZVI), and activated carbon-supported nZVI (A-nZVI), were studied to compare their phytotoxicity in mung bean grown in suspensions with doses of 0, 300, 600 and 1000 mg/L. Taking the four toxicology parameters (seed germination rate, germination index, seedling elongation and biomass) together, the iron-based materials except mFe3O4 generally produced no significant phytotoxicity to mung bean even at 1000 mg/L. nFe3O4 and B-nZVI showed no higher phytotoxicity than their micro-sized counterparts (mFe3O4 and mZVI). All the materials resulted in increased Fe concentrations in seedlings particularly in roots, and mZVI and B-nZVI produced more significant effects. However, the Fe in the roots was difficultly translocated to the shoots. Compared to B-nZVI, nFe3O4 had lower bioavailability and bioaccumulation potential. XRD results confirmed that most Fe3O4 and B-nZVI remained unchanged during seedling growth, while support materials accelerated the corrosion and transformation of S-nZVI and A-nZVI. In conclusion, the tested nanoscale iron-based materials generally possess no obvious phytotoxicity within the dose range, but cause excess Fe accumulation in seedlings. Introduction of support materials may reduce such risk, allowing safer applications of these iron-based materials.

Adsorption characteristics of cadmium onto microplastics from aqueous solutions.

As one of emerging contaminants, microplastics (MPs) can enter the environment and adsorb toxic metals such as cadmium (Cd), thereby causing potential environmental risks. However, adsorption characteristics of MPs are poorly understood. Herein, batch experiments were performed to investigate the adsorption characteristics of Cd onto high-density polyethylene (HDPE) MPs with different particle sizes, that is, 1-2 mm, 0.6-1 mm, and 100-154 µm. The adsorption of Cd was quite rapid initially, and the equilibrium time was approximately 90 min. An increase in the pH of the Cd solution led to an increase in Cd adsorption. MPs with particle size of 100-154 µm had the highest adsorption capacity. Addition of 1, 10, and 100 mg/L NaCl all significantly decreased Cd adsorption. Adsorption kinetics fitted the pseudo-second-order model. Adsorption isotherm followed the Langmuir model and, to a lesser extent, the Freundlich model, with estimated maximum adsorption capacity of 30.5 µg/g. The adsorbed Cd easily desorbed from the MPs. Energy-dispersive X-ray spectroscopy (EDS) analysis confirmed Cd adsorption to and desorption from MPs. Fourier transform infrared (FTIR) spectroscopy analysis showed no new functional groups formed during the adsorption and desorption processes, suggesting physical interaction may dominate the Cd adsorption onto MPs. The present study findings provide evidence that MPs can accumulate Cd, and the adsorbed Cd may be highly available, thus posing risks to the organisms exposed to these MPs.

Evaluating phytotoxicity of bare and starch-stabilized zero-valent iron nanoparticles in mung bean.

Zero-valent iron nanoparticles (nZVI) are among the most widely used nanoparticles in nanoremediation of various environmental pollutants. Environmental fate and impact of nZVI has attracted increasing concerns due to their potential risks. However, phytotoxicity of nZVI still remains poorly understood. Here, the phytotoxic effects of bare nZVI (B-nZVI) and starch-stabilized nZVI (S-nZVI) were evaluated on the germination of mung bean seeds exposed to suspensions with different doses of 0-1000 mg/L and the growth of hydroponically cultured seedling at 600 mg/L. In most cases, B-nZVI had no inhibition on seed germination, and even promotion on shoot and root elongation. However, S-nZVI displayed dose-dependent effects, with a decreased germination rate at 600-750 mg/L. B-nZVI at 600 mg/L showed no obvious phytotoxic but even stimulatory effects on seedling growth. Comparatively, S-nZVI at 600 mg/L produced significant phytotoxicity on mung bean plants, leading to decreased seedling growth, altered nutritional balance, and excess Fe accumulation in roots (>400 mg/kg). S-nZVI were observed to form a coating of insoluble Fe(III) compounds on root surface. Simultaneously, some nZVI penetrated and accumulated into root cells, but did not move to shoots. In conclusion, B-nZVI easily aggregate into larger particles in solution, leading to decreased adhesion to root surface and lower uptake by roots, whereas the higher dispersity and hydrophilicity of S-nZVI makes them more readily be adhered to root surface forming a coating, and penetrated into roots, resulting in excess Fe accumulation, consequently interfering with root functions such as the adsorption and transport of water and nutrients.

Mechanisms and Characterization of Trichoderma longibrachiatum T6 in Suppressing Nematodes (Heterodera avenae) in Wheat.

Heterodera avenae is an important soil-borne pathogen that affects field crops worldwide. Chemical nematicides can be used to control the nematode, but they bring toxicity to the environment and human. Trichoderma longibrachiatum has been shown to have the ability to control H. avenae cysts, but detailed microscopic observations and bioassays are lacking. In this study, we used microscopic observations and bioassays to study the effect of T. longibrachiatum T6 (TL6) on the eggs and second stage juveniles (J2s) of H. avenae, and investigate the role of TL6 in inducing the resistance to H. avenae in wheat seedling at physiological and biochemical levels. Microscopic observations recorded that TL6 parasitized on the H. avenae eggs, germinated, and produced a large number of hyphae on the eggs surface at the initial stage, thereafter, the eggs were completely surrounded by dense mycelia and the contents of eggs were lysed at the late stage. Meanwhile, the conidia suspension of TL6 parasitized on the surface of J2s, produced a large number of hyphae that penetrated the cuticle and caused deformation of the nematodes. TL6 at the concentration of 1.5 × 107 conidia ml-1 had the highest rates of parasitism on eggs and J2s, reflected by the highest hatching-inhibition of eggs and the mortality of J2s. In the greenhouse experiments, wheat seedlings treated with TL6 at 1.5 × 107 conidia ml-1 had reduced H. avenae infection, and increased plant growth significantly compared to the control. The cysts and juveniles in soil were reduced by 89.8 and 92.7%, the juveniles and females in roots were reduced by 88.3 and 91.3%, whereas the activity of chitinase and ß-1, 3-glucanase, total flavonoids and lignin contents in wheat roots were increased significantly at different stage after inoculation with the eggs and TL6 conidia in comparison to the control. Maximum activity of chitinase and ß-1, 3-glucanase were recorded at the 20th and 15th Days after inoculation with TL6 and thereafter it declined. The maximum contents of total flavonoids and lignin were recorded at the 35th and 40th Days after inoculation with TL6. After being stained with the rapid vital dyes of acridine orange (AO) and neutral red (NR), the frozen and infected eggs and J2s of H. avenae changed color to orange and red, respectively, while the color of eggs and J2s in control group did not change. Therefore, our results suggest that TL6 is potentially an effective bio-control agent for H. avenae. The possible mechanisms by which TL6 suppresses H. avenae infection are due to the direct parasitic and lethal effect of TL6 on the eggs and J2s activity, and the induced defense response in wheat plants together.

[Parasitic and lethal action of Trichoderma longibrachiatum against Heterodera avenae].

OBJECTIVE: To evaluate the potential of Trichoderma longibrachiatum spore suspension against Heterodera avenae. METHODS: The parasitic and lethal effects of T. longibrachiatum spore suspension against the cysts of H. avenae were studied in vitro and observed under microscope. RESULTS: Microscopic observation showed that the spore suspension of T. longibrachiatum parasitized on the cyst surface, germinated a large number of hyphae, and grew on the surface of the cyst at the initial stage. Later, the cysts were completely surrounded by dense mycelium, and the contents of digestion in cysts was lysed, even some cysts produced vacuoles, and some were split up and finally the cyst was dissolved by the metabolite of T. longibrachiatum. In vitro studies showed that high concentrations of T. longibrachiatum spores had strong parasitic and lethal effects on the cysts of H. avenae, and the probable mechanism of parasitic and lethal effects of T. longibrachiatum against H. avenae were mainly by inducing and increasing chitinase, glucanase and caseinase activity. The cysts were parasitized by 93.3% at 18 days, the hatching of cysts were inhibited by 93.6% at 10 days when treated with the concentrations (1.5 x 10(8) CFU/mL) of T. longibrachiatum. CONCLUSION: Trichoderma longibrachiatum had strong parasitic and lethal effects on the cysts of H. avenae, and has the potential as a new biocontrol agent.

[Parasitic and lethal effects of Trichoderma longibrachiatum on Heterodera avenae: microscopic observation and bioassay].

A laboratory experiment was conducted to study the parasitic and lethal effects of Trichoderma longibrachiatum conidia suspension on Heterodera avenae cysts. Different concentrations (1.5 x 10(5)-1.5 x 10(7) cfu x mL(-1)) of T. longibrachiatum conidia suspension had strong parasitic and lethal effects on H. avenae cysts, and the effects differed significantly among the different concentrations. When treated with the T. longibrachiatum conidia suspension at a concentration of 1.5 x 10(7) cfu x mL(-1), 96.7% of the H. avenae cysts were parasitized by the conidia at the 18th day, and the hatching rate of the cysts was inhibited by 91.2% at the 22nd day. The microscopic observation showed that at the initial parasitic stage, T. longibrachiatum conidia suspension adhered or parasitized on the cyst surface, germinated a large number of hyphae, and grew on the cyst surface, making the development of cyst embryo stopped and the contents in cysts flocculated, and even, some cysts started to deform, and small dark brown vacuoles formed on the cyst surface. At the later parasitic stage, the cysts were penetrated by dense mycelium, cysts were broken, their contents exosmosed, and the mycelium on the integument of some cysts produced conidiophores, on which, conidium were adhered or parasitized. It was considered that T. longibrachiatum could be used as a potential high-efficient bioagent to control the occurrence and damage of H. avenae.

[Angiogenesis and regulatory factors in rats with BPH induced by testosterone].

OBJECTIVE: To study angiogenesis and regulatory factors in the proliferated prostatic tissues of Sprague Dawley (SD) rats with BPH induced by testosterone. METHODS: Sixteen castrated SD rats, aged 8 weeks and weighing 200 approximately 250 g, were equally randomized into a model group and a control group, and the BPH model was established by subcutaneous injection of testosterone. Immunohistochemistry and MIAS (micro-image analysis system) were used to test the manifestations of MVD (microvessel density), VEGF (vascular endothelium growth factor), flk-1, endostatin, MMP-2 (matrix metalloproteinase-2) and TIMP-2 (tissue inhibitor of metalloproteinase-2) in the prostatic tissues of both the model and the control groups. Multiple linear regression with the stepwise method was adopted to analyze the data. RESULTS: The manifestations of MVD, VEGF, flk-1, MMP-2, MMP-2/TIMP-2 and VEGF/endostatin in the model group were higher, while that of endostatin was lower than in the control group (P < 0.01), and the manifestation of TIMP-2 showed no statistical difference between the two groups. The regression analysis indicated that MVD was positively correlated to VEGF, VEGF/endostatin and MMP-2/TIMP-2 (r = 0.974, 0.986, 0.982, P < 0.05) and negatively correlated to endostatin (r = - 0.975, P < 0.05) . CONCLUSION: Testosterone could induce BPH in SD rats by increasing MVD and promoting the multiplication of vascular endothelial cells after regradation of basement membrane.

[System review of the Chinese medicine bushenhuoxue for treating benign prostatic hyperplasia].

OBJECTIVE: To make a system review of the effects of Bushenhuoxue (kidney-tonifying and blood-activating prescription), a category of compound Chinese medicines, on benign prostatic hyperplasia (BPH) and its side effects. METHODS: All the research articles about Chinese medicines treating BPH from January 1978 to February 2003 were retrieved using the methods of international evidence-based medicine (EBM), and their qualities were evaluated based on JADAD standard and concealment of research allocation. Then the included articles went through META-analysis with Revman 4.2 software. RESULTS: The efficacy of Bushenhuoxue on BPH was better than Qianliekang but not statistically different from finasteride. The combined use of Bushenhuoxue with surgery had no statistical difference from mere surgical treatment. Four articles reported the side effects of this compound Chinese medicine including upset and pain in the abdomen, nausea, diarrhea and dryness of the nose. CONCLUSION: Massive, multi-center and randomized controlled clinical trials should be conducted to find out more effective methods for treating BPH with Chinese medicines based on the improvement of measurable symptom evaluation method and for evaluating their side effects.

[Laboratory research of the influence of keyouling on the proliferation of human prepuce epidermis cells and condyloma acuminatum cells].

OBJECTIVE: To discuss the mechanism of the Chinese medicine Keyouling in the treatment of condyloma axuminatum (CA). METHODS: Human prepuce epidermis cells and CA cells were primarily cultured and subcultured. We determined the proliferation of human prepuce epidermis cells and CA cells, and observed the influence of Keyouling with different concentrations on the proliferation of human prepuce epidermis cells and CA cells by means of MTT colourimetry assay. RESULTS: The absorbance was directly proportional to the numbers of human prepuce epidermis cells (r = 0.9850, P < 0.001) and CA cells (r = 0.9892, P < 0.001). Keyouling had no effect on proliferation of the human prepuce epidermis cells, but it had significant inhibition on CA cells. The concentrations of Keyouling bore negative correlation with the proliferation percentage of CA cells(r = -0.4124, P < 0.01). CONCLUSION: Keyouling can significantly restrain the growth and proliferation of CA cells but has no damaging effect on normal organic cuticle cells. It is suggested that Keyouling might have anti-HPV effect.

[Experimental and clinical research on the effect of keyouling on condyloma acuminatum and adjustment of cellular immunity function].

OBJECTIVE: To discuss the mechanism of the traditional Chinese medicine Keyouling oral liquid in the treatment of condyloma acuminatum(CA) and the adjustment of cellular immunity function. METHODS: The IL-18 and TNF-alpha levels of peripheral serum and wart tissue of patterned rats and CA patients exposed to Keyouling were determined by means of double-antibody sandwich ELISA, and the NK cellular activity of the spleen of the patterned rats and that of the peripheral blood of the CA patients exposed to Keyouling were determined by means of 3H-TdR isotype release. RESULTS: The IL-18 and TNF-alpha levels, the NK cellular activity of the high-dosage group showed significant difference from those of the pattern group and low-dosage group in animal experiment(P < 0.05); the IL-18 and TNF-alpha levels of peripheral serum and wart tissues, and the NK cellular activity of the peripheral blood of the treatment group showed significant difference from those of the control group after treatment(P < 0.01, P < 0.05). CONCLUSIONS: Keyouling oral liquid has significant positive adjusting effect, which can markedly ameliorate the cellular immunadeficiency of the patterned animals and reinforce the cellular immunocompetence of CA patients.

[Experimental and clinical research of "Keyouling" on treatment of condyloma acuminata].

OBJECTIVE: To investigate the effects of traditional Chinese medicine "Keyouling" in treatment of condyloma acuminata (CA). METHODS: The effects of "Keyouling" at different concentrations on growth and proliferation of rabbits' prepuce epithelial cells and CA warts tissue were assessed by means of cellular primary culture and subculture, and the effects of "Keyouling" at different concentrations on the expression level of HPV6.11 DNA and expression level of HPV6.11 DNA of different area of CA patient's penis were quantitatively detected by means of FQ-PCR. RESULTS: "Keyouling" had no effect on growth and proliferation of rabbits' prepuce cuticle cells, but it had significant inhibition on CA warts tissue cells and expression of HPV6.11 DNA. Concentrations of "Keyouling" assumed negative correlated with cell proliferation rate of CA warts tissue and expression level of HPV6.11 DNA, and also had significant inhibition on expression of HPV6.11 DNA of negative cutaneous lesion area and cutaneous lesion area of CA patients. CONCLUSION: "Keyouling" had significant anti HPV6.11 effect, could significantly restrain growth and proliferation of CA warts tissue cells, while it had no damaging effect on normal organic cuticle cells. It was suggested that "Keyouling" was safe in clinical use and had good therapeutic effect on CA and subclinical infection.

[Histomorphological studies on hyperplastic prostate of castrated rat caused by androgen].

OBJECTIVES: To investigate prostatic histomorphological changes of rat benign prostatic hyperplasia (BPH) models. METHODS: After castrated, SD rats were injected subcutaneously testosterone propionate to induce the BPH. Water substitution method was used to measure prostatic volumes. Prostatic tissue were stained by hemoloxylin and eosin, the morphometric changes of glandular and interstitial tissues were semi-quantified by image analysis system. Multiple linear regression was adopted to analyze the results. RESULTS: In comparison with normal group, prostatic volumes were significantly enlarged (P < 0.01) with glands expanded and interstitial tissues increased in BPH model. Glandular average diameters, volumes and surface areas in unit volume, as well as glandular circumferences and glandular relative total volumes were all significantly increased (P < 0.01). Glandular counts, density, ratio of glandular surface area to volume. And glandular average curvature were all declined (P < 0.05-0.01), so was interstitial circumference (P < 0.01). But volume density had no changes, and relative total interstitial volumes were obviously increased. Prostatic volume was significantly correlated with glandular relative total volume (r = 0.989, P < 0.001) and interstitial relative total volume(r = 0.789, P < 0.001). Prostatic volume was also correlated significantly with glandular average volume(r = 0.789, P < 0.001). CONCLUSIONS: Action of androgen on rat prostate may mainly lie in glandular epithelial hyperplasia, which manifests enlargement of glandular lumen accompanied by hyperplasia of interstitial tissues.