Effect of Crop Rotation on Fusarium Mycotoxins and Fusarium Species in Cereals in Sichuan Province (China).

The present study was performed to evaluate the effect of crop rotation on Fusarium mycotoxins and species in cereals in Sichuan Province. A total of 311 cereal samples were randomly collected and analyzed from 2018 to 2019 in Sichuan Province. The results of mycotoxin analysis showed that the major trichothecene mycotoxins in Sichuan Province were nivalenol (NIV) and deoxynivalenol (DON), and the mean concentration of total trichothecenes (including NIV, fusarenone X [4ANIV], DON, 3-acetyldeoxynivalenol [3ADON], and 15-acetyldeoxynivalenol [15ADON]) in wheat was significantly higher than that in maize and rice. The concentration of total trichothecenes in the succeeding crops was significantly higher than that in the previous crops. In addition, wheat grown after maize had reduced incidence and concentration of trichothecene mycotoxins compared with that grown after rice, and ratooning rice grown after rice had increased incidence and concentration of trichothecene mycotoxins. Our data indicated that Fusarium asiaticum with the NIV chemotype was predominant in wheat and rice samples, while the number of the NIV chemotypes of F. asiaticum and Fusarium meridionale and the 15ADON chemotype of Fusarium graminearum in maize were almost the same. Although the composition of Fusarium species was affected by crop rotations, there were no differences when comparing the same crop rotation except for the maize-wheat rotation. Moreover, the same species and chemotype of Fusarium strains originated from different crops in various rotations, but there were no significant differences in pathogenicity in wheat and rice. These results contribute to the knowledge of the effect of crop rotation on Fusarium mycotoxins and species affecting cereals in Sichuan Province, which may lead to improved strategies for control of Fusarium mycotoxins and fungal disease in China.

Network pharmacology and experimental evidence reveal the protective mechanism of Yi-Qi Cong-Ming decoction on age-related hearing loss.

CONTEXT: Yi-Qi Cong-Ming (YQCM) decoction has been widely used to prevent age-related hearing loss (ARHL), the most prevalent neurodegenerative disease in the elderly. OBJECTIVE: To explore the mechanism of YQCM decoction in the treatment of ARHL. MATERIALS AND METHODS: The chemical constituents of YQCM were screened from the Traditional Chinese Medicine Systems Pharmacology Database. Potential targets of YQCM against ARHL were predicted by DrugBank, GeneCards, and OMIM database. Protein-protein network and enrichment analysis were used for exploring possible molecular mechanisms. Molecular docking and an in vitro model of ARHL by exposing auditory cells with 100 µM H2O2 for 3 h were applied. Cell viability and mitochondrial membrane potential (ΔΨM) were detected by CCK-8 and high-content analysis. γH2AX and cleaved caspase-3 were detected by Western blot. RESULTS: The main compounds have good affinities with hub targets, especially AKT1, PTGS2, and CASP3. GO and KEGG analysis showed that the main biological process and key targets were related to negative regulation of the apoptotic process. H2O2 treatment could reduce the cell viability by 68% and impaired ΔΨM, while 90 µg/mL YQCM pre-treatment could restore the cell viability by 97.45% and increase ΔΨM (2-fold higher). YQCM pre-treatment also reduced γH2AX and cleaved caspase-3 protein levels. CONCLUSIONS: Our study suggested that YQCM prevents ARHL by modulating the apoptosis process in auditory hair cells. Moreover, this study proved that bioinformatics analysis combined with molecular docking and cell model is a promising method to explore other possible pharmacological interventions of ARHL.

Analysis of the Fusarium graminearum Species Complex from Gramineous Weeds Near Wheat Fields in Jiangsu Province, China.

Several weed species are known as alternative hosts of the Fusarium graminearum species complex (FGSC), and their epidemiological significance in Fusarium head blight (FHB) has been investigated; however, scant information is available regarding FGSC occurrence in weeds near Chinese wheat fields. To evaluate the potential role of gramineous weeds surrounding wheat fields in FHB, 306 FGSC isolates were obtained from 210 gramineous weed samples in 2018 in Jiangsu Province. Among them, 289 were Fusarium asiaticum, and the remainder were F. graminearum. Trichothecene genotype and mycotoxin analyses revealed that 74.3% of the F. asiaticum isolates were the 3-acetyldeoxynivalenol (3ADON) chemotype, and the remainder were the nivalenol (NIV) chemotype. Additionally, 82.4% of F. graminearum isolates were the 15-acetyldeoxynivalenol (15ADON) chemotype, and the remainder were the NIV chemotype. FHB severity and trichothecene analysis indicated that F. asiaticum isolates with the 3ADON chemotype were more aggressive than those with the NIV chemotype in wheat. 3ADON and NIV chemotypes of F. asiaticum isolated from weeds and wheat showed no significant differences in pathogenicity in wheat. All selected F. asiaticum isolates produced perithecia, with little difference between the 3ADON and NIV chemotypes. These results highlight the epidemiology of the FGSC isolated from weeds near wheat fields, with implications for reducing FHB inoculum in China.

Analysis of Fusarium graminearum Species Complex from Freshly Harvested Rice in Jiangsu Province (China).

Members of Fusarium graminearum species complex (FGSC) are the major pathogens that cause Fusarium head blight (FHB) in cereals worldwide. Symptoms of FHB on rice, including dark staining or browning of rice glumes, were recently observed in Jiangsu Province, China. To improve our understanding of the pathogens involved, 201 FGSC isolates were obtained from freshly harvested rice samples and identified by phylogenetic analyses. Among the 201 FGSC isolates, 196 were F. asiaticum and the remaining 5 were F. graminearum. Trichothecene chemotype and chemical analyses showed that 68.4% of the F. asiaticum isolates were the 3-acetyldeoxynivalenol (3ADON) chemotype and the remainder were the nivalenol (NIV) chemotype. All of the F. graminearum isolates were the 15-acetyldeoxynivalenol chemotype. Pathogenicity assays showed that both the 3ADON and NIV chemotypes of F. asiaticum could infect wheat and rice spikes. FHB severity and trichothecene toxin analysis revealed that F. asiaticum with the NIV chemotype was less aggressive than that with the 3ADON chemotype in wheat, while the NIV-producing strains were more virulent than the 3ADON-producing strains in rice. F. asiaticum isolates with different chemotypes did not show significant differences in mycelial growth, sporulation, conidial dimensions, or perithecial production. These findings would provide useful information for developing management strategies for the control of FHB in China.

Ototoxicity of paclitaxel in rat cochlear organotypic cultures.

Paclitaxel (taxol) is a widely used antineoplastic drug employed alone or in combination to treat many forms of cancer. Paclitaxel blocks microtubule depolymerization thereby stabilizing microtubules and suppressing cell proliferation and other cellular processes. Previous reports indicate that paclitaxel can cause mild to moderate sensorineural hearing loss and some histopathologic changes in the mouse cochlea; however, damage to the neurons and the underlying cell death mechanisms are poorly understood. To evaluate the ototoxicity of paclitaxel in more detail, cochlear organotypic cultures from postnatal day 3 rats were treated with paclitaxel for 24 or 48 h with doses ranging from 1 to 30 µM. No obvious histopathologies were observed after 24h treatment with any of the paclitaxel doses employed, but with 48 h treatment, paclitaxel damaged cochlear hair cells in a dose-dependent manner and also damaged auditory nerve fibers and spiral ganglion neurons (SGN) near the base of the cochlea. TUNEL labeling was negative in the organ of Corti, but positive in SGN with karyorrhexis 48 h after 30 µM paclitaxel treatment. In addition, caspase-6, caspase-8 and caspase-9 labeling was present in SGN treated with 30 µM paclitaxel for 48 h. These results suggest that caspase-dependent apoptotic pathways are involved in paclitaxel-induced damage of SGN, but not hair cells in cochlea.

Phosphodiesterase type 10A inhibitor attenuates lung fibrosis by targeting myofibroblast activation.

Pulmonary fibrosis (PF) is a fatal and irreversible respiratory disease accompanied by excessive fibroblast activation. Previous studies have suggested that cAMP signaling pathway and cGMP-PKG signaling pathway are continuously down-regulated in lung fibrosis, whereas PDE10A has a specifically expression in fibroblasts/myofibroblasts in lung fibrosis. In this study, we demonstrated that overexpression of PDE10A induces myofibroblast differentiation, and papaverine, as a PDE10A inhibitor used for vasodilation, inhibits myofibroblast differentiation in human fibroblasts, Meanwhile, papaverine alleviated bleomycin-induced pulmonary fibrosis and amiodarone-induced oxidative stress, papaverine downregulated VASP/ß-catenin pathway to reduce the myofibroblast differentiation. Our results first demonstrated that papaverine inhibits TGFß1-induced myofibroblast differentiation and lung fibrosis by VASP/ß-catenin pathway.

Plasmoid ejection and secondary current sheet generation from magnetic reconnection in laser-plasma interaction.

Reconnection of the self-generated magnetic fields in laser-plasma interaction was first investigated experimentally by Nilson et al. [Phys. Rev. Lett. 97, 255001 (2006)] by shining two laser pulses a distance apart on a solid target layer. An elongated current sheet (CS) was observed in the plasma between the two laser spots. In order to more closely model magnetotail reconnection, here two side-by-side thin target layers, instead of a single one, are used. It is found that at one end of the elongated CS a fanlike electron outflow region including three well-collimated electron jets appears. The (>1 MeV) tail of the jet energy distribution exhibits a power-law scaling. The enhanced electron acceleration is attributed to the intense inductive electric field in the narrow electron dominated reconnection region, as well as additional acceleration as they are trapped inside the rapidly moving plasmoid formed in and ejected from the CS. The ejection also induces a secondary CS.

[Biological research evaluating the Chinese medical theory of the association of the kidney with the ears].

"Association of the kidney with the ears" is central to the traditional Chinese medical (TCM) theory of the viscera and their manifestations. Previous studies have shown that thyroxin, aldosterone, calcium and iron might be the material basis of the correlation between the kidney and the ears. Due to restriction in research techniques and methods, there has been no further progress in this research field, whose modern biological mechanisms and scientific connotations have not been explained clearly. In recent years, the progress of biological techniques and research on kidney essence provided promising opportunities for research of the association between the kidney and the ears. This article summarized the study progress of "association of the kidney with the ears" theory and posed some meaningful study thoughts. Presbycusis is a kind of degenerative disease caused by aging, which is one aspect of aging of the human body. Because aging is considered as physiological "kidney deficiency", presbycusis can be regarded as a model of "kidney deficiency" deafness. In conclusion, investigating the correlation between "kidney deficiency" and presbycusis using biotechnology systems combined with "essence of kidney", should provide a sound basis for study of the theory of the relationship between the kidney and the ears. This will provide new and valuable information for the modern biological research about the theory of visceral associations in TCM.

[Research progress in biological basis of cold and heat essence of Chinese medicine].

Cold-heat problem is one core of traditional Chinese medicine theory. This paper summarizes the experimental research related to the biological basis of cold-heat essence in cold-heat syndrome, cold-heat body constitution and cold-heat property of Chinese herbs. In view of the classical physiological and biochemical indices, gene expression, protein expression and metabolic differences, differences in cold-heat syndrome or cold-heat constitution are mainly based on neurotransmitter, thyroid function, sex hormone, cyclic nucleotide system, and energy metabolism relating to the corresponding gene and protein expression. Furthermore, this paper analyses the change of correlation indices that accompany with a dynamic development process of "constitution-syndrome-herbal intervention", implying that the research of biological basis of cold-heat essence has turned from single index to multiple indices, and from dispersion research to system research.

Synthetic antimicrobial agents inhibit aflatoxin production.

Antimicrobial peptides (AMPs) are biologically active molecules that can eradicate bacteria by destroying the bacterial membrane structure, causing the bacteria to rupture. However, little is known about the extent and effect of AMPs on filamentous fungi. In this study, we synthesized small molecular polypeptides by an inexpensive heat conjugation approach and examined their effects on the growth of Aspergillus flavus and its secondary metabolism. The antimicrobial agents significantly inhibited aflatoxin production, conidiation, and sclerotia formation in A. flavus. Furthermore, we found that the expression of aflatoxin structural genes was significantly inhibited, and the intracellular reactive oxygen species (ROS) level was reduced. Additionally, the antimicrobial agents can change membrane permeability. Overall, our results demonstrated that antimicrobial agents, safe to mammalian cells, have an obvious impact on aflatoxin production, which indicated that antimicrobial agents may be adopted as a new generation of potential agents for controlling aflatoxin contamination.

Transient receptor potential A1 is involved in cold-induced contraction in the isolated rat colon smooth muscle.

Transient receptor potential (TRP) A1, a member of TRP channel family, is activated by noxious cold. The aims of this study were to determine if TRPA1 contributed to cold-induced contractions in the isolated rat colon preparations and explore the potential mechanisms. The colon smooth muscle layers were surgically isolated from the male Wistar rats and changes in isotonic tension of longitudinal muscle under various treatments were recorded as colonic motilities. Cold stimuli were obtained by the reperfusion with Krebs-Henseleit solution at given temperature using Constant Flow Pump. The mRNA expressions of TRPA1, TRPV1 and TRPM8 in rat colon smooth muscle layer were examined by using reverse transcription-polymerase chain reaction (RT-PCR) techniques. The results showed that the contractions induced by cold stimuli (from 37 degrees C to 12 degrees C stepwise) were inversely proportional to the temperature with a maximum contraction at 17 degrees C in both proximal and distal colons (P<0.01). RT-PCR analysis revealed the expression of TRPA1, but not TRPM8 and TRPV1, in the rat proximal and distal colon smooth muscle layers. Cold-induced colonic contractions were specially inhibited by TRPA1 blocker, ruthenium red (30 µmol/L), in the proximal and distal colon (P<0.05). The cold-induced contractions of proximal (P<0.01, P<0.05) and distal colons (both P<0.001) were almost abolished or inhibited by the pretreatments of TRPA1 agonists, Allyl isothiocyanate (AITC, 300 µmol/L) and cinnamaldehyde (CA, 1 mmol/L). Extracellular calcium removal (EGTA, 1 mmol/L), PLC blocker (U73122, 10 µmol/L) and IP(3) receptor blocker (2-aminoethoxydiphenyl borate, 2-APB, 30 µmol/L) all decreased the contractions evoked by the cooling at 17 degrees C in the proximal and distal colon (P<0.001, P<0.05, P<0.001). Atropine (1 µmol/L) had no effects on these contractions. L-type Ca(2+) channels blocker nifedipine (1 µmol/L) and neurotoxin tetrodotoxin (TTX, 2 µmol/L) decreased the contractile response in the distal colon (P<0.01, P<0.05), but not in the proximal colon. In conclusion, TRPA1 contributes to cold-induced contractions of the rat colon smooth muscle, and the mechanism of TRPA1 activation involves PLC/IP(3)/Ca(2+) pathway. L-type Ca(2+) channel and neurogenic mechanism other than muscarinic receptor might be partially involved in cold-induced contraction of the distal colon, which probably resulted in higher contraction of distal colon compared with that of proximal colon.

[Effect of electrical stimulation of the primary auditory cortex on the spontaneous activities of the external nucleus of the inferior colliculus in a rat model of tinnitus induced by salicylate acid.].

This study aims to investigate corticofugal modulation on spontaneous activities of the external nucleus of the inferior colliculus (ICx) in a salicylate acid (SA) induced tinnitus rat model by the stimulation of the primary auditory cortex (AI). Extracellular recording techniques and stereotaxic method were used. The spontaneous activities of a single unit were recorded from the left ICx after electrical stimulation was given to the left AI of the rats duplicated as acute SA models. The average rate of spontaneous discharge and the interspike interval histogram of spontaneous activities were used as indices. The single unit spontaneous discharges of the same unit of ICx before and after AI stimulation were observed. There was an inhibitory effect of AI stimulation on the activities of the high discharge unit [(8.75+/-2.70) Hz vs (5.06+/-2.01) Hz] and a facilitatory effect on the low discharge unit [(1.41+/-0.45) Hz vs (2.46+/-0.79) Hz]. In the normal group, there was a restraining effect on the average rate of spontaneous discharge of the ICx after AI stimulation. The average rate of spontaneous discharge changed from (3.66+/-0.84) Hz to (2.47+/-0.43) Hz in the first hour after AI stimulation and then recovered within 2-4 h. And the discharge rate of the spike interval within 0-20 ms decreased (17% vs 7.3%, 11.2%) in the first 2 h and recovered 3-4 h after AI stimulation. The discharge rate of the spike interval within 0-6 ms (short interval) recovered 2 h after AI stimulation. In the acute SA model group, the average rate of spontaneous discharge recorded from the ICx decreased from (7.48+/-0.85) Hz to (3.38+/-0.39) Hz in the first hour after AI stimulation and the suppression effect remained 4 h (P<0.05). There was no difference in the average rate of spontaneous discharge between the acute SA model group and the normal group at 2-4 h after AI stimulation. The suppression effect after AI stimulation on the 0-20 ms interval spikes in the ICx lasted 4 h, while that on the shorter interval (0-6 ms) spikes recovered in the 3rd hour after AI stimulation in acute SA model group. It is concluded that the high average spontaneous discharge rate of ICx in acute SA model decreases significantly by AI stimulation, and the suppression effects on the shorter interval spikes recovers in the 3rd hour after AI stimulation. It might be inferred that stimulation of AI, through exciting the auditory descending projections, could remit the increased spontaneous discharge of ICx induced by SA that may relate with tinnitus in a period of time.

[Cloning and expression analysis of two salt and Fusarium graminearum stress associated UDP-glucosyltransferases genes in wheat].

Glycosyltransferases (GTs) play important roles in stress responses of plants by glycosylating hormones and secondary metabolites. UDP-glucosyltransferases (UGTs), which use UDP-glucuronic acid in animals, UDP-glucose, UDP-galactose, and UDP- rhamnose in plant as sugar donors, belong to family 1 of GTs. As a secondary metabolite produced by Fusarium graminearum during infection of grains, deoxynivalenol (DON) is not only harmful to human and animal's health by inhibiting protein synthesis, but also acts as a virulence factor during fungal pathogenesis. In order to study expression profile of UGT genes in wheat, two UGTs, designated TaUGT1 and TaUGT2, were isolated from bread wheat (Triticum aestivum L.) by reverse transcriptase-polymerase chain reaction. The genomic sequences of both genes had one intron. Their coding sequences shared 91% and 90% similarities at nucleic acid level and the deduced protein sequence level. The analysis of conserved domain revealed that these two cDNAs encoded UDP-glucoronosyl and UDP-glucosyl transferase with PSPG (Putative secondary plant glycosyltransferase) domain. Real-time PCR was carried out to detect the expression profiles of the two UGTs in wheat under various stress conditions. In young spikes infected by Fusarium graminearum, TaUGT2 was induced but TaUGT1 was repressed. These two genes were upregulated under higher NaCl concentration. In conclusion, TaUGT2 may participate in wheat resistance to Fusarium head blight in which mycotoxin DON plays a role, and these two genes might be involved in responses of wheat to salt stress.

Association between age­related hearing loss and cognitive decline in C57BL/6J mice.

Accumulating evidence has revealed the link between age­related hearing loss (presbycusis) and cognitive decline; however, their exact association remains unclear. The present study aimed to investigate the association between age­related hearing loss and cognitive decline, and to explore the underlying mechanisms. Briefly, three groups of C57BL/6J mice were evaluated, based on their age, as follows: Young group, 3 months; adult group, 6 months; and middle­aged group, 15 months. The results of an auditory brainstem response (ABR) test demonstrated that the hearing threshold levels of the mice were increased in those aged 6 and 15 months compared with those aged 3 months, thus suggesting that significant hearing loss occurred at 6 months, and worsened at 15 months. The results of a Morris water maze test demonstrated that spatial learning and memory function was significantly decreased in 15­month­old mice, but not in 6­month­old mice. Pearson analysis indicated that the escape latency was positively correlated with hearing threshold at 16 kHz and percentage of time in the target quadrant was negatively correlated with hearing threshold at 16 kHz, thus suggesting a correlation between age­related hearing loss and cognitive decline. The auditory cortex and hippocampal CA1 region in 15­month­old mice exhibited significantly decreased cell numbers, abnormal arrangement and morphological alterations. Transmission electron microscopy revealed reduced synapse numbers and synaptic vesicle density in mice aged 15 months. Furthermore, the protein expression levels of matrix metalloproteinase (MMP)­9 in the auditory cortex and hippocampus in the 15­month­old mice were significantly higher than in the 3­month­old mice. In conclusion, these findings support the correlation between age­related hearing loss and cognitive decline in C57BL/6J mice, and indicated that MMP­9 expression in the auditory cortex and hippocampus may be associated with the underlying mechanisms.

[Effects of Erxian Decoction and its separate prescriptions on the levels of luteinizing hormone and follicle-stimulating hormone in anterior pituitary cells from female rats].

OBJECTIVE: To investigate the effects of Erxian Decoction (EXD), a traditional Chinese compound herbal medicine and its separate prescriptions such as Wenshen Yijing Recipe (WSYJR, a recipe for warming kidney and replenishing essence), Ziyin Xiehuo Recipe (ZYXHR, a recipe for nourishing yin and dispersing fire) and Tiaoli Chongren Recipe (TLCRR, a recipe for regulating thoroughfare and conception vessels) and some extracts from EXD on the levels of LH and FSH in the primary cultured anterior pituitary cells from female rats. METHODS: EXD, its separate prescriptions and traditional Chinese herbal extracts from EXD were added directly to the incubators. The levels of LH and FSH were tested by radioimmunoassay. RESULTS: The levels of LH and FSH in the supernatant of anterior pituitary cells treated by EXD or its separate prescriptions including ZYXHR and TLCRR were increased significantly as compared with those in the blank control. There was a tendency for stimulating the secretions of LH and FSH in the WSYJR-treated group too. Dimethyl sulfoxide as the solvent of icariin and curculigoside could interfere with the results. CONCLUSION: EXD and its separate prescriptions such as ZYXHR and TLCRR can increase the levels of LH and FSH significantly.

[Effect of er xian decoction and its subdivisions on granulosa cells secretory function in rats].

OBJECTIVE: To investigate the effects of Er Xian Decoction (EXD) and its three subdivisions "warming Shen" ,"nourshing Yin" and "adjusting Chong and Ren" in regulating the level of estradiol (E2) on the primary cultured granulosa cells. METHODS: Effect of EXD and its three subdivisions, also part of the effective components of this formula, icariin and curculigoside, on level of E2 were carried out using primary cultural granulose cell as the experimental model. RESULT: EXD and its three subdivisions could stimulate the secretion of E2, especially the "warming Shen" group (P <0.05). All the composing of Chinese herbs of this formula could promote the level of E2 in different degree, and the Rhizoma Curculiginis, Radix Moromade Officinalis, and Herba Epimedii have the best effects (P <0. 01). CONCLUSION: The regulation of EXD on the hypothalamus-pituitary-gonad (HPG) axis may be related to promoting the secreting of E2 at the site of granulosa cell. The "warming Shen" subdivision has the better effect in promoting the secretion of E2.

Temporal dynamics, population characterization and mycotoxins accumulation of Fusarium graminearum in Eastern China.

Trichothecene genotype composition, mycotoxin production, genetic diversity, and population structure were analyzed, using 185 Fusarium strains collected from wheat (Triticum aestivum L.) throughout the Jiangsu province during 1976, 1983, 1998, 2006, and 2014. The results showed that 3-acetyldeoxynivalenol (3ADON) was consistently the predominant type in this region over 40 years, and the nivalenol (NIV) type has emerged since 1998. Long-term rotation of wheat and rice (Oryza sativa L.), rather than fungicide application, crop fitness, or weather conditions, might be the main cause of this phenomenon. The genetic diversity results from two toxin synthetic genes, Pks4 and Tri10, and variable number of tandem repeat (VNTR) markers revealed the largest variance within the population in 1998, which was also the year with the highest production of mycotoxins. Population differentiation analysis indicated that major temporal population comparisons from the same area were not significantly differentiated. Our results showed that dominant species could maintain genetic stability for a long time, and Pks4 would be of utility in genetic and population studies.

Effects of Erlong Zuoci decoction on the age-related hearing loss in C57BL/6J mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Erlong Zuoci decoction (ELZCD), a typical traditional Chinese medicine (TCM) prescription, has long been clinically used in treatment of deafness and tinnitus with the syndrome of "kidney yin deficiency". However, there are few studies to investigate its pharmacological mechanisms. Until now, there is not report about its effects on the age-related hearing loss (ARHL). AIM OF STUDY: The present study was conducted to observe the effects of ELZCD on the ARHL in C57BL/6J mice and explore the mechanisms. MATERIALS AND METHODS: ELZCD was fed to C57BL/6J mice from 3 months to 6 months in ELZCD group as a dose of 6g/kg/d. And the same volume of saline was fed to mice in ARHL group. 3-months-old C57BL/6J mice were used as control group. High performance liquid chromatography (HPLC) was used for the quality control of ELZCD. Auditory brainstem response (ABR) was used to assess the hearing function of mice. The morphologic changes were observed by hematoxylin eosin (HE) staining. Apoptosis was tested by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) method. Mitochondrial damage was detected by transmission electron microscopy (TEM). Quantitative RT-PCR (qRT-PCR) was used to observe the mRNA expression of p53 and Bak. Fluorescence immunohistochemical technique was used to test the protein expression of p53 and Bak. RESULTS: The hearing threshold of ARHL group was higher than that of control group (P<0.001) and ELZCD decreased the rise of hearing threshold levels of ARHL mice (P<0.001), which suggested ELZCD inhibited the hearing loss of ARHL mice. HE staining showed that ELZCD decreased the spiral ganglion (SG) cell damage and loss in ARHL. TUNEL test showed that the apoptotic SG cells increased in ARHL group compared to control group and decreased in ELZCD group compared to ARHL group. TEM observation showed that mitochondrial damage was obvious in SG cells of ARHL group and ELZCD inhibited the mitochondrial damage. The qRT-PCR results showed that the mRNA expression of p53 and Bak in ARHL group increased compared to that of control group (P<0.05), and ELZCD reduced the elevated mRNA expression levels of p53 and Bak (P<0.01, P<0.05). In addition, ELZCD inhibited the increased proteins expression (green fluorescence) of p53 and Bak. CONCLUSION: The results demonstrated that ELZCD prevented ARHL in C57BL/6J mice and p53/Bak-mediated mitochondrial apoptosis of SG cells might be involved in the mechanisms.

[Effects on secretory function of rat gonad by Erxian decoction and its disassembled prescriptions].

OBJECTIVE: To explain functions, differences and coordination of three divided combinations of the "Erxian decoction", the famous traditional Chinese formula, on the effective sites of gonad gland at the cell level. METHOD: The effects of Erxian decoction and its main disassembled prescriptions, "Kidney Warming", "Yin Nourishing" and "Chong-ren Adjusting", on the level of testosterone (T) progesterone (P) estradiol (E2), respectively secreted by the primary culture Leydig cell, luteal cell and granulosa cell, were measured by radioimmunoassay. RESULT: (1) Erxian decoction could stimulate the T secretion while its three main disassembled prescriptions would seem no individual promoting effect on the secretion of T. (2) Erxian decoction and the "Kidney Warming" had the stimulating effect on P secretion, and the action of the whole formula being better than that of the "Kidney Warming". (3) Erxian decoction and its main disassembled prescriptions had the stimulating effect on E2 secretion, especially the whole formula. CONCLUSION: Erxian decoction can stimulate the secretion of T of the Leydig cell, P of luteal cell and E2 of granulosa cell. It can be seen that the effect of the whole formula is better than that of its main disassembled prescriptions.