Drynaria Naringin alleviated mechanical stress deficiency-caused bone loss deterioration via Rspo1/Lgr4-mediated Wnt/ß-catenin signalling pathway.

Osteoporosis is a metabolic condition distinguished by the degradation of bone microstructure and mechanical characteristics. Traditional Chinese medicine (TCM) has been employed in China for the treatment of various illnesses. Naringin, an ingredient found in Drynariae TCM, is known to have a significant impact on bone metabolism. For this research, we studied the precise potential effect of Drynaria Naringin on protecting against bone loss caused by stress deficiency. In this study, a tail-suspension (TS) test was performed to establish a mouse model with hind leg bone loss. Some mice received subcutaneous injections of Drynaria Naringin for 30 d. Trabecular bone microarchitecture was evaluated using micro-computed tomography analysis and bone histological analysis. Bone formation and resorption markers were quantified in blood samples from mice or in the supernatant of MC3T3-E1 cells by ELISA analysis, Western blotting, and PCR. Immunofluorescence was utilized to visualize the location of ß-catenin. Additionally, siRNA was employed to knockdown-specific genes in the cells. Our findings highlight the efficacy of Drynaria Naringin in protecting against the deterioration of bone loss and promoting bone formation and Rspo1 expression in a mouse model following the TS test. Specifically, in vitro experiments also indicated that Drynaria Naringin may promote osteogenesis through the Wnt/ß-catenin signalling pathway. Moreover, our results suggest that Drynaria Naringin upregulates the expression of Rspo1/Lgr4, leading to the promotion of osteogenesis via the Wnt/ß-catenin signalling pathway. Therefore, Drynaria Naringin holds potential as a therapeutic medication for osteoporosis. Drynaria Naringin alleviates bone loss deterioration caused by mechanical stress deficiency through the Rspo1/Lgr4-mediated Wnt/ß-catenin signalling pathway.

Ethanol extract of Ilex hainanensis Merr. exhibits anti-melanoma activity by induction of G1/S cell-cycle arrest and apoptosis.

OBJECTIVE: To evaluate anti-melanoma effect of ethanol extract of Ilex hainanensis Merr. (IME) and elucidate its underlying mechanism. METHODS: Thirty-six tumor-bearing mice were randomized into 6 groups (n=6) as follows: model group, IME 25, 50, 100, and 200 mg/kg groups and dacarbazine (DTIC) 70 mg/kg group. The mice in the IME treatment groups were intragastrically administered with IME 25, 50, 100 or 200 mg/kg per day, respectively. The mice in the DTIC group were intraperitoneally injected with DTIC 70 mg/kg every 2 days. The drug administration was lasting for 14 days. The cell viability was evaluated by 3-(4,5-dime-thylthylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Flow cytometry was employed to detect cell cycle and apoptosis. The gene and protein expressions of nuclear factor κB-p65 (NF-κB-p65), Bcl-2, B-cell lymphomaextra large (Bcl-xL) and Bax were detected by quantitative real-time polymerase chain reaction and Western blot analyses. Caspases-3, -8, and -9 activities were detected using the colorimetric method. In addition, a B16-F10 melanoma xenograft mouse model was used to evaluate the anti-cancer activity of IME in vivo. Furthermore, a survival experiment of tumor-bearing mice was also performed to evaluate the possible toxicity of IME. RESULTS: IME significantly inhibited the proliferation of B16-F10 cells (P<0.01). Flow cytometric analysis showed that IME induced G1/S cell cycle arrest and apoptosis (both P<0.01). IME inhibited activation of NF-κB, decreased the gene and protein expressions of Bcl-2, Bcl-xL, and increased the gene and protein expressions of Bax (all P<0.01). In addition, IME induced the activation of Caspases-3, -8, and -9 in B16-F10 cells. The study in vivo showed that IME significantly reduced tumor volume (P<0.01), and the inhibitory rate came up to 68.62%. IME also induced large areas of necrosis and intra-tumoral apoptosis that correlated with a reduction in tumor volume. Survival experiment showed that treatment with IME for 14 days significantly prolonged survival time and 20% of mice in the IME 200 mg/kg group were still alive until the 50th day. Notably, IME showed no apparent side-effects during the treatment period. CONCLUSION: IME exhibited significant anti-melanoma activity in vitro and in vivo, suggesting that IME might be a promising effective candidate with lower toxic for malignant melanoma therapy.

Ilexgenin A induces B16-F10 melanoma cell G1/S arrest in vitro and reduces tumor growth in vivo.

The present study aimed to investigate the anti-tumor activity of Ilexgenin A in B16-F10 murine melanoma and to evaluate its effect on the production of tumor-associated inflammatory cytokines. In vitro, our study showed that Ilexgenin A inhibited the proliferation of B16-F10 murine melanoma cells in a dose- and time-dependent manner, and this effect could be ascribed to the arrest of the cell cycle at G0/G1. In vivo, we evaluated the anti-tumor activity of Ilexgenin A in a tumor-bearing mouse model. The results showed that Ilexgenin A reduced the tumor weight by 51.13% (p<0.01). The Ilexgenin A treatment groups showed no apparent side effects during the treatment period. In addition, a histological analysis revealed that Ilexgenin A changed the cell morphology, and induced large areas of necrosis that correlated with a reduction in tumor size. The detection of inflammatory cytokines indicated that the IL-6 level decreased (p<0.001) and the TNF-α level increased (p<0.01) in mice treated with Ilexgenin A. Ilexgenin A also inhibited the IL-6 production of macrophages stimulated by melanoma conditioned medium (MCM) significantly (p<0.001). Importantly, Ilexgenin A dramatically prolonged survival time (p<0.001). In conclusion, Ilexgenin A could be regarded as a promising agent for the treatment of melanoma; it exerts anti-melanoma activity by arresting the cell cycle at G0/G1 and regulating IL-6 and TNF-α production.

Clinical treatment of the stenosing tenovaginitis of flexor digitorum by micro-wound technique using hooked needle-shaped surgical knife.

OBJECTIVE: To observe the therapeutic effect and safety of using the hooked needle-shaped knife to treat the stenosing tenovaginitis of flexor digitorum. METHODS: Sixty outpatients were divided into a treatment group of 30 cases treated by using the hooked needle-shaped knife, and a control group of 30 cases treated by block therapy. 6 months later, the alleviation of pain in the affected finger during movement, under pressure, traction and finger-bending anti-resistance was compared before and after treatment. RESULTS: The alleviation of pain during movement, under pressure, traction and finger-bending anti-resistance was much better in the treatment group than that of the control group (P < 0.01), with a effective rate of 93.3% in the treatment group and 80.0% in the control group (P < 0.01). CONCLUSION: The micro-wound technique using the hooked needle-shaped knife has definite effect and safety for stenosing tenovaginitis of flexor digitorum.

The chemical composition and ultrastructure of uroliths in Boer goats.

The chemical composition and ultrastructure of urinary calculi obtained from male Boer goats were studied using qualitative chemical analysis, scanning electron microscopy, X-ray diffraction, X-ray energy dispersive spectrometry and Fourier transform infra-red spectroscopy. The calculi came from 10 naturally-occurring cases of urolithiasis and from seven cases of urolithiasis experimentally-induced by feeding a cottonseed meal-rice straw diet supplemented with magnesium oxide. The results indicated that the major component of urinary calculi collected from naturally-occurring and experimentally-induced cases of urolithiasis was struvite (magnesium ammonium phosphate). The study also identified previously unreported prismatic crystals in the uroliths of goats, similar to struvite but rich in potassium. The characteristic ultrastructure of struvite uroliths is described along with a brief discussion of their formation.

Study on precipitation of struvite and struvite-K crystal in goats during onset of urolithiasis.

To learn more about the biomineralization process of struvite in ruminants, a seldom noticeable crystal, struvite-K, was investigated in six goats in which urolithiasis was induced by feeding a cottonseed meal and rice straw diet supplemented with MgO. The compositions of crystals and calculi were studied by chemical qualitative analysis and X-ray energy dispersive spectrometry (EDS). The activity product (AP) and relative supersaturation (RSS) of magnesium ammonium phosphate (MAP) and potassium magnesium phosphate (MKP) were calculated to compare the difference of crystals formed in this process. The results showed that calculi consisted of MAP with a little MKP. Crystals in the urine consisted of MAP and MKP before stone formation, but crystals in urine after stone formation mainly consisted of MKP. The AP and RSS of MAP and MKP significantly decreased after stone formation. It was concluded that MAP and MKP may coexist in the crystals of urine before struvite calculi formation but MKP did not precipitate to struvite calculi and was separated out as crystal sedimentation of urine. The changes of crystallization of MAP and MKP contributed partially to the supersaturation status of MAP and MKP during struvite stone formation.

[Case control study of hook needle knife for the treatment of stenosing tenovaginitis of flexor digitorum].

OBJECTIVE: To investigate the effect and safety of the hook needle knife for the treatment of stenosing tenovaginitis of flexor digitorum. METHODS: From September 2007 to September 2008, 60 outpatients with stenosing tenovaginitis of flexor digitorum were randomized divided into the treatment group and the control group, 30 cases in each group. Among the patients, 44 patients were female and 16 patients were male, aged from 34 to 69 years, averaged 56 years, the duration of disease ranged from 1 month to 1 year, averaged 3 months. All the patients were treated with hook needle knife and local-blocking respectively. The patients were followed up for 6 months, and the relief of moving-pain, tender-pain, stretching-pain and resist-ing--pain were observed respectively. All the patients were evaluated by the symptoms with numerical rating scale. RESULTS: The relief of moving-pain, tender-pain, stretching-pain and resisting-pain in the treatment group were significantly better than those of the control group; and the therapeutic effects of treatment group were better than those of the control group. CONCLUSION: The method for treating stenosing tenovaginitis of flexor digitorum with hook needle knife has advantages of definite effects, micro-invasion and safety.

Comparison of effect of high intake of magnesium with high intake of phosphorus and potassium on urolithiasis in goats fed with cottonseed meal diet.

The effect of high intake of Mg on urolithiasis was compared with high intake of P and K in goats being fed with a cottonseed meal and rice straw diet. Eighteen wether goats were randomly allocated into group A, B and C evenly and fed with cottonseed meal and rice straw diet for three months. From day 60 onwards, KH(2)PO(4) and K(2)HPO(4) were provided via drinking water to goats in group B to increase the intake of P, K, and MgO to goats in group C to increase the intake of Mg. Blood and urine samples were collected to analyze the concentration of P, K, Mg and Ca, and the activity product (AP) of potassium magnesium phosphate (MKP) in urine was also calculated. The composition of calculi and urinary sedimentary crystals were examined by chemical qualitative analysis, X-ray diffraction, X-ray energy dispersive spectrometry and Fourier transform infrared spectroscopy. The results showed that the incidence of urolithiasis in group C (6/6) was higher than that in group A (1/6) and B (1/6) (P<0.05). The calculi were mainly composed of magnesium ammonium phosphate (MAP) and partly composed of MKP. MKP presented in crystals of different phases in this experiment. The high intake of Mg contributed to a significant increase of plasma Mg, but additional P, K did not cause a further increase of plasma P, K. Urine P, K, Mg and Ca and AP of MKP in group C decreased significantly after the onset of urolithiasis. In conclusion, high intake of Mg was more important in inducing struvite calculi compared with high intake of K and P in goats under these feeding conditions. Cottonseed meal and rice straw with additional Mg is a good dietary model for inducing struvite calculi in castrated goats.

L-arginine prevents reduced expression of endothelial nitric oxide synthase (NOS) in pulmonary arterioles of broilers exposed to cool temperatures.

This study investigated nitric oxide synthase (NOS) expression in the endothelium of pulmonary arterioles of broilers during the development of pulmonary hypertension syndrome (PHS). PHS was triggered by exposing broilers to sub-thermoneutral (cool) temperatures and an additional 1.0% L-arginine was added to the basal diet to evaluate the effects of supplemental L-arginine on nitric oxide (NO) production, endothelial NOS expression, and the incidence of PHS. Cumulative mortality from PHS, right/total ventricle weight ratios (RV/TV), and body weights were recorded. Plasma NO concentration and NOS expression in the endothelium of pulmonary arterioles with an outer diameter ranging from 100 to 200 microm were determined. Birds exposed to cool temperatures had increased pulmonary hypertension and PHS mortality and diminished endothelial NOS expression. Supplemental dietary L-arginine reduced PHS mortality and elicited higher NOS expression within the pulmonary endothelium coincident with elevated NO production. The results demonstrated that broilers developing PHS exhibited diminished NOS expression in the endothelium of their pulmonary arterioles. Supplemental L-arginine prevented the reduced expression of NOS in the pulmonary endothelium, which might contribute to the increased production of NO by the pulmonary vasculature.

L-Arginine inhibiting pulmonary vascular remodelling is associated with promotion of apoptosis in pulmonary arterioles smooth muscle cells in broilers.

OBJECTIVE: Pulmonary vascular remodelling is one of the important pathological bases of broiler pulmonary hypertension syndrome (PHS). Nitric oxide (NO) has been found to inhibit proliferation and to induce apoptosis in pulmonary artery smooth muscle cells (SMC) in mammals with pulmonary hypertension. The present study was conducted to evaluate the effects of NO precursor l-arginine on pulmonary vascular remodelling in broilers with pulmonary hypertension induced by cold exposure and to examine whether NO-induced apoptosis in pulmonary arteriole SMC is involved in the regulatory mechanisms. METHODS: Two hundred and forty mixed-sex commercial broilers were equally assigned to three groups and reared in normal brooding temperatures before day 14. Starting on day 14 continuing until the end of the experiment, the control group was brooded in normal temperatures whereas the other two groups were subjected to low ambient temperatures with or without l-arginine added to the basal diets. Cumulative PHS mortality and body weight were recorded in each group. Right/total ventricle ratio (RV/TV), plasma NO concentration and pulmonary vascular morphological changes were analyzed. TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay was used to detect apoptosis in pulmonary arteriole SMC. RESULT: l-Arginine, in group A, had no effect on body weights under cold temperature condition. Birds kept in group B had increased PHS mortality, RV/TV ratio, vessel wall area/vessel total area ratios (WA/TA) and mean media thickness in pulmonary arterioles (mMTPA) (P<0.05). Percentages of apoptotic SMC in pulmonary arterioles in group B were not altered by cold exposure (P>0.05). Supplemental dietary l-arginine in group A elevated plasma NO level (P<0.05), reduced PHS mortality (P<0.05), attenuated pulmonary vascular remodelling and increased the percentages of apoptotic SMC (P<0.05) when compared with the group B. CONCLUSION: Supplemental l-arginine partially inhibited pulmonary vascular remodelling that occurred secondary to increased pulmonary pressure; NO-induced apoptosis in arteriole SMC might contribute to its regulatory effect on pulmonary vascular structural changes.