Triptolide inhibits the proinflammatory potential of myeloid-derived suppressor cells via reducing Arginase-1 in rheumatoid arthritis.

Triptolide (TPT) is widely used in the treatment of rheumatoid arthritis (RA). However, its regulatory mechanisms are not fully understood. This study demonstrated that Myeloid-derived suppressor cells (MDSCs) were expanded in both RA patients and arthritic mice. The frequency of MDSCs was correlated with RA disease severity and T helper 17 (Th17) responses. MDSCs from RA patients promoted the polarization of Th17 cells in vitro, which could be substantially attenuated by blocking arginase-1 (Arg-1). TPT inhibited the differentiation of MDSCs, particularly the monocytic MDSCs (M-MDSCs) subsets, as well as the expression of Arg-1 in a dose dependent manner. Alongside, TPT treatment reduced the potential of MDSCs to promote the polarization of IL-17+ T cell in vitro. Consistently, TPT immunotherapy alleviated adjuvant-induced arthritis (AIA) in a mice model, and reduced the frequency of MDSCs, M-MDSCs and IL-17+ T cells simultaneously. The presented data suggest a pathogenic role of MDSCs in RA and may function as a novel and effective therapeutic target for TPT in RA.

A novel method for making cell blocks with higher cellular yield.

INTRODUCTION: Cytopathology is an important part of pathology that is used to diagnose disease on the cellular level. The application of the cell block (CB) technique plays a vital role in cytological diagnosis, as blocks and slides can be further used for special stains, immunohistochemistry (IHC), and molecular pathological analysis. Several methods for making CBs have been reported, but their procedures and cellular yield are still deemed unsatisfactory. In this article, we used gellan gum (GG) as an adjuvant for CBs, which resulted in higher cellular yield with simpler procedures. METHODS: CBs were prepared by using GG, copper sulfate, plasma/thrombin, or pregelatinized starch methods. The procedures of each of these four methods were then compared. CB sections were stained with hematoxylin and eosin (H&E), and the background and morphological features seen by H&E staining were compared. A preliminary IHC and fluorescence in situ hybridization (FISH) study was performed using cytology specimens from eleven and five cases, respectively. The expression of immunocomplex by IHC and the molecular signals detected by FISH were compared in CB sections made by the four methods and a section derived from the biopsy specimen block from the same patient. Feulgen staining, Alcian blue staining, and Masson trichrome staining were performed on the CB sections from 3 cases of pleural fluid. The cellular yield of CB sections from 83 cases according to the four methods was compared using NDP analysis software. RESULTS: The results demonstrated that sections derived from CBs made with GG had a clear background and good morphological features by H&E staining. The expression of immunocomplex by IHC and the molecular signals of FISH detection in the sections from CBs made by GG were accurately located just as those in biopsy sections from the same patient. The DNA, acidic mucus, and fibrin could be clearly identified through special stains in the CB sections. The procedures involved in the GG method were easily controllable and the coagulated gel increased the ease by which the CB was embedded and sectioned. Specifically, sections from CBs made by the GG method contained higher cellular yield because cells could be concentrated on the bottom of the gel after centrifugation. CONCLUSION: This novel method for making CBs is a practical, simple method that can result in higher cellular yield. This method is therefore worth promoting in clinical applications.

Radix astragali inhibits the down-regulation of connexin 26 in the stria vascularis of the guinea pig cochlea after acoustic trauma.

Connexin 26 (cx26) plays an important role in the intercellular signaling and is related to K(+) metabolism in stria vascularis (SV). Reactive oxygen species (ROS) are negative regulators of cx26, reducing intercellular coupling in cochlea. ROS plays an important role in acoustic trauma. Radix astragali is a natural antioxidant that decreases impulse noise-induced hearing loss through its ability to inhibit ROS. The purpose of this study was to investigate if radix astragali has the potential to reduce the change of cx26 in SV from impulse noise. Guinea pigs in the experimental group were administered radix astragali intraperitoneally. Auditory thresholds were assessed by sound-evoked auditory brainstem response (ABR) at click and tone bursts of 8, 16 and 32 kHz, 24 h before and 72 h after exposure to impulse noise. 4-Hydroxynonenal, cx26 and KCNQ1 were determined immunohistochemically in SV. SV was analyzed by transmission electron microscopy. Radix astragali significantly reduced the ABR deficits and the SV damage, and decreased the shifts of the expression of cx26 and KCNQ1 in the SV. These results suggest that the beneficial effect of radix astragali on impulse noise-induced hearing loss may be also due to its ability to reduce the change of cx26 in SV.

Radix astragali injection enhances recovery from sudden deafness.

OBJECTIVES: An acute interruption of the blood supply to the inner ear is one of the most likely causative factors for sudden deafness (SD). Reactive oxygen species (ROS) have been suggested to be important mediators of the tissue injury during cochlear ischemia and reperfusion. Radix astragali (RA) is natural antioxidant. The aim of this study was to investigate the efficacy of RA in patients with SD. PATIENTS AND METHODS: We compared the hearing gains from hearing impairment in 46 ears treated with RA with 46 ears treated with non-RA. RA was given intravenously daily for 10 days. There were no significant differences in clinical or audiological data between RA and non-RA groups. RESULTS: The hearing gain at 250, 500, 1000, 2000, and 4000 Hz in RA group was much higher than that of non-RA group correspondingly (P < .01). Also, the hearing gain at PTA (pure-tone average of 250, 500, 1000, 2000, and 4000 Hz) in RA group was significantly higher than that of non-RA group (P < .01). CONCLUSION: The recovery of hearing was significantly better after treatment of RA than non-treatment of RA. RA can be valuable concurrent therapy for patients with SD.

Astragaloside IV inhibits apoptotic cell death in the guinea pig cochlea exposed to impulse noise.

CONCLUSION: The results suggest that the beneficial effect of astragaloside IV on impulse noise-induced hearing loss may be due to its ability to inhibit reactive oxygen species (ROS) and prevent apoptosis. OBJECTIVE: Astragaloside IV is the major active constituent of Astragalus membranaceus, which has been widely used for the treatment of diseases in China for its antioxidant properties. ROS and apoptosis are involved in damage induced by impulse noise trauma. We aimed to investigate if the beneficial effects of astragaloside IV on cochlea exposed to impulse noise are associated with the inhibition of ROS and the decrease in apoptosis. METHODS: 4-Hydroxynonenal (HNE) was used as the marker of ROS. Active-caspase-3 (cas-3) served as a marker for apoptosis. 4HNE and cas-3 were determined immunohistochemically. Guinea pigs in the experimental group were administered astragaloside IV intragastrically. Auditory thresholds were assessed by sound-evoked auditory brainstem response (ABR) 72 h before and after exposure to impulse noise. RESULTS: The results showed that astragaloside IV significantly reduced ABR deficits, and decreased the expression of ROS and cas-3.

Radix Astragali injection enhances recovery from acute acoustic trauma.

CONCLUSION: The average recovery of hearing and cessation of tinnitus was significantly better after treatment with Radix Astragali (RA) than after non-treatment with RA. RA can be valuable adjuvant therapy for patients with acute acoustic trauma (AAT). OBJECTIVES: AAT is one of the early indications for the use of RA. The reasons for administering RA to patients with AAT are based on experimental studies showing that noise exposure results in the formation of reactive oxygen species (ROS), which trigger metabolic damage to the organ of Corti. RA is a natural antioxidant. The aim of this study was to investigate the efficacy of RA in patients with AAT. METHODS: We compared the recovery from hearing impairment and tinnitus in 40 ears treated with RA with 40 ears treated with non-RA. RA was given intravenously daily for 10 days. There were no significant differences in clinical or audiological data between RA and non-RA groups. RESULTS: The average recovery of hearing at both high and speech frequencies was significantly better and tinnitus persisted less commonly in the RA group than in the non-RA group. Normal hearing at the end of the follow-up period was regained in 27 ears in the RA group and in 21 ears in the non-RA group (p < 0.01).

Oxidative stress in spiral ganglion cells of pigmented and albino guinea pigs exposed to impulse noise.

CONCLUSIONS: The results suggest that melanin inhibits formation of reactive oxygen species (ROS) and prevents apoptosis in spiral ganglion cells (SGCs) of pigmented guinea pigs following impulse noise. OBJECTIVE: The stria vascularis of pigmented guinea pig cochlea contains melanocytes that produce melanin, which has a protective effect on noise-induced hair cell damage through its antioxidant property. ROS are involved in cochlear damage induced by impulse noise trauma. The purpose of the present study was to investigate the oxidative stress in SGCs of pigmented and albino guinea pigs after exposure to impulse noise. METHODS: Pigmented and albino guinea pigs were exposed to impulse noise. Auditory thresholds were assessed by sound-evoked auditory brainstem response (ABR) before impulse noise exposure and 72 h after impulse noise exposure. 4-Hydroxynonenal (HNE) was used as a histochemical marker of ROS formation, and active-caspase-3 (cas-3) served as a marker for apoptosis. 4-HNE and cas-3 were determined immunohistochemically. Hair cell damage was analyzed by scanning electron microscopy. RESULTS: The rates of 4-HNE-positive and cas-3-positive SGCs in pigmented guinea pigs were much less than those for albino guinea pigs. Correspondingly, there was less hair cell damage and reduced ABR threshold shifts in pigmented guinea pigs.

Astragalosides reduce cisplatin ototoxicity in guinea pigs.

Cisplatin is known to cause high-frequency neurosensory hearing loss. While reactive oxygen species have been shown to play a role, reactive nitrogen species have been implicated, but not proven to be involved, in cisplatin ototoxicity. The purpose of the present study was to investigate the role of nitric oxide (NO) in cisplatin ototoxicity by administering astragalosides, a natural antioxidant, in conjunction with cisplatin. Guinea pigs were injected with cisplatin, astragalosides or both. Auditory brainstem-evoked responses (ABRs) were measured before and 3 days after cisplatin administration. The cochlear tissue was then assayed for NO and malondialdehyde (MDA), and cochleae were also examined by scanning electron microscopy. Cisplatin alone caused significant ABR threshold shifts at all stimuli tested, whereas astragalosides alone caused no shifts. There was a significant reduction in threshold shift for clicks, 8-kHz and 16-kHz tone bursts (but not 32 kHz) when astragalosides was given with cisplatin. Both the MDA concentration and the NO concentration in the astragalosides/cisplatin group were significantly lower than those of the cisplatin group. Correspondingly, the loss of outer hair cells in the astragalosides/cisplatin group was much less than that in the cisplatin group. This suggests that astragalosides reduces cisplatin ototoxicity by its antioxidant property.

Astragaloside IV attenuates impulse noise-induced trauma in guinea pig.

CONCLUSION: These results suggest that the beneficial effect of astragaloside IV on impulse noise-induced hearing loss may be due to its ability to inhibit inducible nitric oxide synthase (iNOS) and prevent the formation of reactive nitrogen species (RNS). OBJECTIVE: Astragaloside IV is the major active constituent of Astragalus membranaceus, which has been widely used for the treatment of diseases in China due to its antioxidant properties. iNOS and RNS are involved in damage induced by impulse noise trauma. The purpose of the present study was to investigate if astragaloside IV has the potential to reduce cochlear damage from impulse noise. METHODS: Guinea pigs in the experimental group were administered astragaloside IV intragastrically. Auditory thresholds were assessed by sound-evoked auditory brainstem response (ABR) at click and tone bursts of 8, 16 and 32 kHz, 72 h before and after exposure to impulse noise. iNOS and nitrotyrosine were determined immunohistochemically. Hair cell damage was analyzed by scanning electron microscopy. RESULTS: Astragaloside IV significantly reduced ABR deficits, reduced hair cell damage, and decreased the expression of iNOS and RNS formation.