Vitexin exerts protective effects against calcium oxalate crystal-induced kidney pyroptosis in vivo and in vitro.

BACKGROUND: Nephrolithiasis is a common urinary disease with a high recurrence rate of secondary stone formation. Several mechanisms are involved in the onset and recurrence of nephrolithiasis, e.g., oxidative stress, inflammation, apoptosis, and epithelial-mesenchymal transition (EMT). Vitexin, a flavonoid monomer derived from medicinal plants that exert many biological effects including anti-inflammatory and anticancer effects, has not been investigated in nephrolithiasis studies. Moreover, pyroptosis, a form of programmed cell death resulting from inflammasome-associated caspase activation, has not been studied in mice with nephrolithiasis. PURPOSE: We aimed to investigate the protective effect and underlying mechanisms of vitexin in nephrolithiasis, and the related role of pyroptosis in vivo and in vitro. METHODS: Mouse models of nephrolithiasis were established via intraperitoneal injection of glyoxylate, and cell models of tubular epithelial cells and macrophages were established using calcium oxalate monohydrate (COM). Crystal deposition and kidney tissue injury were evaluated by hematoxylin and eosin, and von Kossa staining. Renal oxidative stress indexes including malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and catalase (CAT), were analyzed. The renal expression of interleukin-1 beta (IL-1ß), gasdermin D (GSDMD), osteopontin (OPN), CD44, and monocyte chemotactic protein 1 (MCP-1), and EMT-related proteins in renal tubular epithelial cells was assessed. Cell viability and the apoptosis ratio were evaluated. RESULTS: In vivo, vitexin alleviated crystal deposition and kidney tissue injury, and decreased the level of MDA, and increased the levels of SOD, GSH, and CAT. Vitexin also reduced the levels of the pyroptosis-related proteins GSDMD, NLRP3, cleaved caspase-1, and mature IL-1ß, which were elevated in mice with nephrolithiasis, and repressed apoptosis and the expression of OPN and CD44. Moreover, vitexin mitigated F4/80-positive macrophage infiltration and MCP-1 expression in the kidneys. Furthermore, an in vitro study showed that vitexin increased the viability of HK-2 cells and THP-1-derived macrophages, which was impaired by treatment with COM crystals, decreased the medium lactate dehydrogenase (LDH) level, and inhibited the expression of pyroptosis-related proteins in HK-2 cells and macrophages. Vitexin repressed EMT of HK-2 cells, with increased expression of pan-cytokeratin (Pan-ck) and decreased expression of Vimentin and alpha-smooth muscle actin (α-SMA), and downregulated the Wnt/ß-catenin pathway. Moreover, vitexin suppressed tumor necrosis factor-α (TNF-α) and IL-1ß mRNA expression, which was upregulated by COM in macrophages. CONCLUSION: Vitexin exerts protective effects against nephrolithiasis by inhibiting pyroptosis activation, apoptosis, EMT, and macrophage infiltration. In addition, GSDMD-related pyroptosis mediates nephrolithiasis.

Downregulation of inflammatory mediators by ethanolic extract of Bergenia ligulata (Wall.) in oxalate injured renal epithelial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: In the Indian traditional system of medicine, Bergenia ligulata (Wall.) Engl. has been used for treatment of urolithiasis. Its efficacious nature has led to its incorporation in various commercial herbal formulations such as Cystone and Neeri which are prescribed for kidney related ailments. AIM OF THE STUDY: To assess whether ethanolic extract of B. ligulata can mitigate the cascade of inflammatory responses that cause oxidative stress and ultimately cell death in renal epithelial cells exposed to hyperoxaluric conditions. MATERIAL AND METHODS: Bioactivity guided fractionation using solvents of varying polarities was employed to evaluate the potential of the extracts of B. ligulata to inhibit the crystallization process. Modulation of crystal morphology was visualized through Scanning electron microscopy (SEM) analysis. Cell death was assessed using flow cytometry based assays. Alteration in the inflammatory mediators was evaluated using real time PCR and immunocytochemistry. Phytochemical characterization of the ethanolic extract was carried out using FTIR, LC-MS and GC-MS. RESULTS: Bioactivity guided fractionation for the assessment of antilithiatic activity revealed dose dependent inhibition of nucleation and aggregation process of calcium oxalate crystals in the presence of various extracts, however ethanolic extract showed maximum inhibition and was chosen for further experiments. Studies on renal epithelial NRK-52E cells showed, cytoprotective efficacy of B. ligulata extract against oxalate injury. SEM anaysis further revealed the potential of the extract to modulate the crystal structure and adhesion to renal cell surface. Exposure of the renal cells to the extract led to conversion of the calcium oxalate monohydrate (COM) crystals to the less injurious calcium oxalate dihydrate (COD) form. Expression analysis for oxidative stress and inflammatory biomarkers in NRK-52E cells revealed up-regulation of Mitogen activated protein kinase (MAPK), Osteopontin (OPN) and Nuclear factor- ĸB (NF-ĸB), in response to calcium oxalate insult; which was drastically reduced in the presence of B. ligulata extract. Flow cytometric evaluation pointed to caspase 3 mediated apoptotic cell death in oxalate injured cells, which was attenuated by B. ligulata extract. CONCLUSION: Considering the complex multifactorial etiology of urolithiasis, ethanolic extract from B. ligulata can be a promising option for the management of kidney stones, as it has the potential to limit inflammation and the subsequent cell death.

Fu-Fang-Jin-Qian-Cao herbal granules protect against the calcium oxalate-induced renal EMT by inhibiting the TGF-ß/smad pathway.

CONTEXT: Nephrolithiasis is a major public health problem worldwide and Fu-Fang-Jin-Qian-Cao granules (FFJQC) is a traditional Chinese herbal formula that is used to treat nephrolithiasis. The main component of nephrolithiasis is calcium oxalate (CaOx) and the epithelial-mesenchymal transition (EMT) shown to play a crucial role in CaOx-induced kidney injury. However, the mechanism underlying the therapeutic effect of FFJQC on the CaOx-induced renal EMT is unknown. OBJECTIVE: This study explores the therapeutic benefits and mechanism of FFJQC in oxalate-induced kidney injury. MATERIALS AND METHODS: 60 male C57BL/6 mice were used in this experiment and divided into 6 groups. A mouse kidney stone model was created by intraperitoneal injection of glyoxylate at a dose of 100 mg/kg for 6 days. The standardized FFJQC was used to treat mouse crystal kidney injury by gavage at 1.35 and 2.7 g/kg, respectively. Western blotting and immunostaining for E-cadherin, cytokeratin 18 (CK18), vimentin, smooth muscle α-actin (α-SMA) and transforming growth factor ß (TGF-ß)/Smad pathway were conducted on renal tissues. RESULTS: Following CaOx-induced kidney injury, the levels of E-cadherin and CK18 in kidney decreased, while vimentin and α-SMA levels increased. The FFJQC treatment increased the levels of E-cadherin and CK18 and decreased vimentin and α-SMA levels in varying degrees. What's more, the FFJQC reduced the expression of CaOx-induced fibrosis marker collagen II. CONCLUSION: FFJQC alleviated the CaOx-induced renal EMT and fibrosis by regulating TGF-ß/smad pathway. Therefore, the FFJQC is an important traditional Chinese medicine for the treatment of CaOx-induced renal injury and fibrosis.

The mechanism of the preventive effect of Shen'an capsule on the calcium oxalate crystal-induced early renal injury based on metabolomics.

Kidney stone disease is a worldwide metabolism-associated disorder with a high incidence of renal dysfunction. However, effective methods to prevent crystalline nephropathy are still lacking owing to the absence of aetiological research. Shen'an (SA) capsules are prepared from Chinese medicinal compounds and are considered a promising treatment for the prevention of crystal-induced renal injury. In this study, 24 mice were randomly divided into four groups: saline, oxalate, SA-treated (via preventive administration) and SA-only groups. A metabolomics analysis based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was performed to explore the plasma metabolic profiles among the different groups. The amount of crystal deposition and the decline in kidney function were significantly alleviated by the use of SA capsule. A total of 24 metabolites that showed a reversal trend following SA capsule administration were identified as plasma biomarkerss of the preventive effects of SA capsules on crystal-induced renal injury. Most of these metabolites were involved in the metabolisms of lipid metabolism, energy metabolism, glutathione metabolism and vitamin metabolism. In conclusion, SA capsules exert a preventive effect in mice with crystal-induced kidney injury via the regulation of multiple metabolic pathways.

Targeting of regulated necrosis in kidney disease.

The term acute tubular necrosis was thought to represent a misnomer derived from morphological studies of human necropsies and necrosis was thought to represent an unregulated passive form of cell death which was not amenable to therapeutic manipulation. Recent advances have improved our understanding of cell death in acute kidney injury. First, apoptosis results in cell loss, but does not trigger an inflammatory response. However, clumsy attempts at interfering with apoptosis (e.g. certain caspase inhibitors) may trigger necrosis and, thus, inflammation-mediated kidney injury. Second, and most revolutionary, the concept of regulated necrosis emerged. Several modalities of regulated necrosis were described, such as necroptosis, ferroptosis, pyroptosis and mitochondria permeability transition regulated necrosis. Similar to apoptosis, regulated necrosis is modulated by specific molecules that behave as therapeutic targets. Contrary to apoptosis, regulated necrosis may be extremely pro-inflammatory and, importantly for kidney transplantation, immunogenic. Furthermore, regulated necrosis may trigger synchronized necrosis, in which all cells within a given tubule die in a synchronized manner. We now review the different modalities of regulated necrosis, the evidence for a role in diverse forms of kidney injury and the new opportunities for therapeutic intervention.

Protective Effects of Pistacia lentiscus L. fruit extract against calcium oxalate monohydrate induced proximal tubular injury.

ETHNOPHARMACOLOGICAL RELEVANCE: The world prevalence of kidney stones is increasing and plants are frequently used to treat urolithiasis. Pistacia lentiscus L, a plant which freely grows around the Mediterranean basin areas, is widely used for various pathologies. P. lentiscus has an important impact as it has economical value on top of its pharmacological interest. Decoctions of its aerial parts and/or resin are used to treat kidney stones. AIM OF THE STUDY: To in vitro assess the potential nephroprotective effect of Pistacia lentiscus ethanolic fruit extract (PLEF) on proximal tubular cells in response to the adhesion of calcium oxalate monohydrate (COM) crystals. MATERIALS AND METHODS: Human Kidney [HK]-2 cells were incubated with and without COM in the presence or absence of PLEF. Cell viability was measured by the resazurin assay. The expression of E-cadherin was analyzed by PCR. The extracellular production of H2O2 was measured by Amplex® Red H2O2 Assay. The numbers of detached or non-adherent COM crystals in the presence of PLEF were microscopically captured and counted using ImageJ software. The interaction of PLEF with COM and the effect of PLEF on crystal size were analyzed by flow cytometry. The spectrophotometric measurement of turbidity was performed for assessing the COM concentration. RESULTS: PLEF incubated with COM was able to increase the cell viability. The decrease of E-cadherin expression after incubation with COM was counteracted by PLEF. Overproduction of H2O2 induced by COM was also inhibited by PLEF. Observations using flow cytometry showed that interactions between PLEF and the COM crystals occurred. PLEF was also effective in reducing the particles size and in lowering COM concentration. CONCLUSION: Our data show that COM tubulotoxicity can be significantly reversed by PLEF -at least in part- via an inhibition of COM crystals adhesion onto the apical membrane. This early beneficial effect of PLEF needs to be further investigated as a useful strategy in nephrolithiasis prevention.

Aspectos botânicos e clínicos das intoxicações por plantas das Famílias Araceae, Euphorbiaceae e Solanaceae no Estado de Pernambuco / Bothanical and clinical aspectos of intoxications caused by plants of the Araceae, Euphorbiaceae and Solanaceae family in the state of Pernambuco

Trata-se de um estudo investigativo sobre os aspectos botânicos e clínicos das intoxicações humanas provocadas por espécies das famílias Araceae, Euphorbiaceae e Solanaceae. É um estudo transversal, descritivo, com abordagem quantitativa, aprovado pelo Comitê de Ética do Hospital da Restauração. Os dados referentes ao período de 1992 a 2009 foram coletados no Centro de Assistência Toxicológica de Pernambuco (CEATOX). Dos 214 prontuários analisados, 140 tiveram diagnóstico de intoxicação por espécies das famílias Araceae (55%; n=77); Euphorbiaceae (36,43%; n=51) e Solanaceae (8,57%; n=12). A maioria das Araceae foi representada por Dieffenbachia amoena Bull (50%); Euphorbiaceae por Manihot esculenta Crantz (11,42%) e Solanaceae por Brugmansia suaveolens (Willd.) Bercht. & J. Presl. (5,71%), utilizadas como ornamental, alimentícia, medicinal, em brincadeiras infantis e suicídio. Em consequência da ingestão de partes dessas plantas os sintomas apresentados foram: edema (língua, lábio), náusea, diarreia, rubor facial, midríase, alucinações e dores abdominais. O tratamento constou de observação clínica (45,31%) e tratamento sintomático (40,18%). A gravidade das intoxicações foi classificada como aguda moderada em 79,69% dos pacientes.(AU)

This is an investigative study about the clinical and botanical aspects of human poisoning caused by plants of the species Araceae, Euphorbiaceae and Solanaceae. It is a cross-sectional descriptive study with a quantitative approach, approved by the Restoration Hospital Ethics Committee. Data was collected at the Toxicological Assistance Centre of Pernambuco (CEATOX) comprising the period of 1992 to 2009. 214 records were analyzed, 140 had an intoxication diagnostic from the families: Araceae (55%, n = 77); Euphorbiaceae (36,43%; n = 51) and solanaceae (8,57%; n = 12). Aracea was majorly represented by Dieffenbachia Amoena Bull (50%); Euphorbiaceae by Manihot esculenta Crantz (11,42%) and Solanaceae by Brugmansia suaveolens (Willd.) Bercht. & J. Presl. (5, 71%) that were used as ornamental plants, food, medicine, in children's play and suicide attempts. As a result of ingestion of parts of the plant the symptoms were edema (tongue, lips), nausea, diarrhea, facial flushing, mydriasis, hallucinations and abdominal pain. Treatment consisted of clinical observation (45,31%) and symptomatic treatment (40,18%). The severity of the intoxications was classified as 'moderate acute' in 79,69% of patients.(AU)

Isolation and prevention of calcium oxalate-induced apoptotic death and oxidative stress in MDCK cells by diosgenin.

Calcium oxalate monohydrate (COM) has been shown to be the most frequent constituent of kidney stones. The interactions of cells with COM crystals produce a variety of physiological and pathological changes including the development of oxidative stress, cellular injury and apoptosis. On the other hand, diosgenin, a steroidal sapogenin, is well known for its antioxidant activity. Therefore, the aim of this study was to evaluate whether diosgenin protects MDCK renal epithelial cells from COM-induced apoptotic death. Diosgenin was isolated from fruits of Solanum xanthocarpum by silica gel column chromatography. It was obtained in high yields (1.23%) and the purity was ascertained by HPTLC analysis. Characterization of diosgenin was done by mp, UV-visible spectrophotometry, elemental analysis, FT-IR, (1)H NMR and (13)C NMR analysis. Cells were co-incubated with COM (80µg/cm(2)) and diosgenin (2.5, 5, 7.5 and 10µg/mL) for 24h. It was found that diosgenin attenuated the apoptotic death induced by COM as measured in terms of cell viability, caspase -9/3 activities and DNA fragmentation percent. The inhibitory role of diosgenin on caspase -9/3 activities was also analyzed using molecular docking experiments, which showed interactions to their active sites by H-bonds. Diosgenin also attenuated the increase in lipid peroxidation and glutathione depletion induced by COM crystals. In conclusion, the preventive effect of diosgenin is associated to the inhibition of oxidative stress and caspases.

[Comparisons of crystal form of raphides to toxicity raphides in four poisonous herbs of Araceae family].

OBJECTIVE: To compare the crystal form with the toxicity intensitity of raphides in four poisonous herbs of Araceae family. METHOD: The four kinds of raphides were extracted and isolated from Pinellia ternate, P. pedatisecta, Arisaema amurense and Typhonium giganteum. These raphides were observed with scanning electron microscopy (SEM), and the elements were analyzed with X-ray photoelectron spectroscopy. Infrared spectrum was used for detecting the functional groups. Toxic intensities of the four kinds of raphides were detected by mice acute toxicity experiment, and the value of LD0 were from caculated by Bliss rule. RESULT: The raphides in the four plants have the similar crystal form. Observation with SEM showed a pointed and blunt end, and a long groove and barbs on a raphide. The raphides in P. ternate and P. pedatisecta were sharper than that in other two, respectively. The results of X-ray diffraction, photoelectric spectra showed that the major component of raphides was calcium oxalate monohydrate, and also showed the elements of N and S existing. Infrared spectra showed the raphides contained functional groups of -COOH and -NH2. These results illustrated that the calcium oxalate monohydrate was not the only component of the raphide. The raphides could produce severe toxic reactions. LD50 values of P. ternate, P. pedatisecta, A. amurense and T. giganteum were 14.78, 14.11, 16.02 and 18.90 mg x kg(-1) (ip), respectively. The corresponding LD50 values of crude drugs were all above 3000 mg x kg(-1) (ip). The toxicity of raphides was 200 times of crude drugs'. CONCLUSION: The raphides in P. ternate and P. pedatisecta, A. amurense and T. giganteum were their common poisonous factor.

Proteomic analysis of calcium oxalate monohydrate crystal-induced cytotoxicity in distal renal tubular cells.

Calcium oxalate monohydrate (COM) is the major crystalline component found in kidney stones and its adhesion to renal tubular cells provokes tubular injury, which in turn enhances COM crystal adhesion. However, COM-induced toxic effects in these tubular cells remain largely unknown. We performed a proteomics study to characterize changes in the cellular proteome in MDCK distal renal tubular cells after an exposure to high-dose (1000 microg/mL) COM crystals for 48 h, at which percentage of cell death was significantly increased. Proteins were extracted from MDCK cells cultured with COM-containing or COM-free medium ( n = 5 individual flasks per group), resolved in individual 2-D gels, and stained with SYPRO Ruby fluorescence dye. Quantitative and statistical analyses revealed 53 proteins whose abundance levels were altered (25 were increased, whereas other 28 were decreased) by COM-induced toxicity. Among these, 50 were successfully identified by quadrupole time-of-flight (Q-TOF) mass spectrometry (MS) and/or tandem MS (MS/MS) analyses. The proteomic data were clearly confirmed by 2-D Western blot analysis. While three chaperones (GRP78, Orp150 and Hsp60) were increased, other proteins involved in protein biosynthesis, ATP synthesis, cell cycle regulator, cellular structure, and signal transduction were decreased. These data provide some novel mechanistic insights into the molecular mechanisms of COM crystal-induced tubular toxicity.

[Study on processing for Rhizoma Pinelliae Praeparatum by comprehensive evaluation of multiple mark].

OBJECTIVE: To investigate the best processing technology of Rhizoma Pinelliae Praeparatum. METHODS: L9(3(4)) orthogonal design was used with four factors: quantities of quick lime and Glycyrrhiza urallensis, processing time and temperature, the contents of Calcium oxalate crystals, Guanosine and Glycyrrhizic acid were determined by RP-HPLC, the irritation test was detected by swollen rabbits eyes with processing products. RESULTS: The optimized processing technology was satisfied with some conditions as follows: the processing time was 48 hours, the processing temperature was 30 degrees C, quantities of quick lime and Glycyrrhiza uralensis were 10 g and 15 g. CONCLUSION: The optimized processing method by orthogonal design has achieved the goal to reduce toxicity, the processing time and the processing temperature are confirmed respectively, and the processing time is significantly shortened comparing with Pharmacopoeia.

[Study on processing mechanism of Pinellia ternate].

OBJECTIVE: Elucidating the detoxification mechanism of the raw Pinellia ternata processed by alum solution or alkaline solution (pH > 12). METHOD: Raw Pinellia ternata was immersed in alum solution and alkaline solution according to Chinese pharmacopoeia. Observed the shape's changing of needle-like calcium oxalate crystals by scanning electro-microscopy. Determinating the contents of calcium oxalate crystals by applying oxidation reduction titration. Measured the irritations of raw P. ternata and various processing products on the model of rabbits'eyes. RESULT: After processed by 8% alum solution prescribed in Chinese pharmacopoeia or 10% sodium carbonate solution, the needle-like shape of raphides in raw Pinellia ternata were changing and the sting barb of raphides were rusted and dissolved, the contents of calcium oxalate crystal in raw Pinellia ternata were sharply declined from more than 1% to less than 0.5%. the decline of contents is relative to the irritation decline of P. ternata on rabbit's eyes. Less than 0.5% calcium oxalate crystals of P. ternata almost had no irritation. CONCLUSION: After processed by 8% alum solution or sodium carbonate solution (pH > 12) , the irritation components in raw P. ternata could be rusted and dissolved, the needle point of raphides was broken, which led to the raphides'content declining and the irritation disappearing. The micro-structures, shapes and contents of calcium oxalate crystals in different medicine plants were not same. These properties of calcium oxalate crystal in India Madder Root and yam et al were very different from those in raw P. ternata.

Dietary oxalate and calcium oxalate nephrolithiasis.

PURPOSE: Patients with calcium oxalate kidney stones are advised to decrease the consumption of foods that contain oxalate. We hypothesized that a cutback in dietary oxalate would lead to a decrease in the urinary excretion of oxalate and decreased stone recurrence. We tested the hypothesis in an animal model of calcium oxalate nephrolithiasis. MATERIALS AND METHODS: Hydroxy-L-proline (5%), a precursor of oxalate found in collagenous foods, was given with rat chow to male Sprague-Dawley rats. After 42 days rats in group 1 continued on hydroxy-L-proline, while those in group 2 were given chow without added hydroxy-L-proline for the next 21 days. Food and water consumption as well as weight were monitored regularly. Once weekly urine was collected and analyzed for creatinine, calcium, oxalate, lactate dehydrogenase, 8-isoprostane and H(2)O(2). Urinary pH and crystalluria were monitored. Rats were sacrificed at 28, 42 and 63 days, respectively. Renal tissue was examined for crystal deposition by light microscopy. RESULTS: Rats receiving hydroxy-L-proline showed hyperoxaluria, calcium oxalate crystalluria and nephrolithiasis, and by day 42 all contained renal calcium oxalate crystal deposits. Urinary excretion of lactate dehydrogenase, 8-isoprostane and H(2)O(2) increased significantly. After hydroxy-L-proline was discontinued in group 2 there was a significant decrease in urinary oxalate, 8-isoprostane and H(2)O(2). Half of the group 2 rats appeared to be crystal-free. CONCLUSIONS: Dietary sources of oxalate can induce hyperoxaluria and crystal deposition in the kidneys with associated degradation in renal biology. Eliminating oxalate from the diet decreases not only urinary oxalate, but also calcium oxalate crystal deposits in the kidneys and improves their function.

Calcium oxalate crystals and methyl salicylate as toxic principles of the fresh leaves from Palicourea longiflora, an endemic species in the Amazonas state.

The species of the genus Palicourea (Rubiaceae family) is well-known for its toxicity towards animals, particularly livestock. This work reports the occurrence of skin irritation during the manipulation of Palicourea longiflora, considering the prevalence of the monofluoracetic acid (MFAA) and another toxic compound: methyl salicylate. The MFAA was identified by 19F-NMR and methyl salicylate by gas chromatography linked to mass spectrometry (GC/MS) analysis. Additionally, an anatomical study of leaves had been used to explain the mechanism of penetration of the toxic principles.

[Study on irritation of calcium oxalate crystal in raw Pinellia ternata].

OBJECTIVE: Confirm the irritation of needle-like calcium oxalate crystals in raw Pinellia ternata. METHOD: Comparing the irritations of raw P. ternate containing calcium oxalate crystals, the raw P. ternate no containing calcium oxalate crystals, the pure needle-like calcium oxalate crystals isolated from raw P. ternata, the extracts of water and various solvents from raw P. ternate. by using the model of rabbits' eyes. Studying the quantity effect relationship of different concentration suspensions of needle-like calcium oxalate crystal isolated from raw P. ternate on rabbits' eyes. Observing the shape and appearance of calcium oxalate crystals in raw P. ternate and raw India Madder Root by the electro microscope and comparing their irritation degrees with the same contents of calcium oxalate crystals. RESULT: Calcium oxalate crystals in raw P. ternata showed very strong irritation property. Under the same content of calcium oxalate crystals, the irritation of raw P. ternata and pure needle-like calcium oxalate crystals isolated from raw P. ternate had no significant difference. The concentrations of needle-like calcium oxalate crystals were do relative to the degree of irritation on rabbits' eyes and they showed undoubted quantity-effect relationship. CONCLUSION: Calcium oxalate crystal is the irritation component in raw P. ternata.

Effect of aqueous extract from Herniaria hirsuta L. on experimentally nephrolithiasic rats.

Despite considerable progress in medical therapy, there is no satisfactory drug to treat kidney stones. Therefore, this current study is aimed to look for an alternative treatment by using Herniaria hirsuta on nephrolithiasic rats as a preventive agent against the development of kidney stones. The experiment was conducted in normal and calcium oxalate (CaOx) nephrolithiasic rats during 3 weeks. Several parameters were followed weekly including water intake, urinary volume and pH, some urinary chemistries, and crystalluria. At the end, kidneys were analyzed by light microscope. The results showed that water intake and urinary volume increased in nephrolithiasic rats, but their urinary pH decreased especially in the third week of treatment. Urinary oxalate increased significantly during the second week for untreated rats and remained constant in rats treated with Herniaria decoction. However, urinary calcium decreased significantly in week 2 in untreated rats and remained constant in treated rats. Qualitative analysis of crystalluria showed that untreated rats excreted large CaOx monohydrate and few dihydrate crystals while treated animals excreted mostly small CaOx dihydrate crystals. The examination of kidney sections revealed that CaOx deposition was limited in treated rats when compared to untreated ones. These results obtained in vivo confirmed the beneficial effect of Herniaria hirsuta and may justify its use as a preventive agent against the formation of calcium oxalate kidney stones.

[Study of stinging crystals in tian nanxing].

OBJECTIVE: To test whether the raphides in Tian Nanxing (Pinellia pedatisecta Schott) caused irritation. METHOD: Scan Electron Microscope and Microscope Oberservation; Animal experimental study. RESULT: Through the comparision of unprocessed Tian Nanxing to processed ones which included 36 h, 72 h, 120 h processed samples, the great modifications in the structure of raphides, especially for the ones with barbs in the processed samples was observed with time course study. A further animal experimental study went to show that the rate of change for raphides with barbs existed a dose-reponse relationship to irritation. CONCLUSION: Investigations of the causes of these reactions showed that raphides of calcium oxalate are, at least in part, responsible for the Tian Nanxing's irritation.

[Studies on stimulating components of raw Pinellia ternata (Thunb.) (banxia)].

OBJECTIVE: To study the stimulating components of raw Pinellia ternata (Banxia). METHODS: Used the methods of extracting with various organic solvents; x-ray fluorescence spectrometer; scanning electron microanalyzer; oxidation and reduction titrations; and ultraviolet spectrophotometry. RESULTS: Experiments showed that calcium oxalate hydrate (C2CaO4 x H2O) was one of the stimulating components of raw Pinellia ternata, and its shape, and contents related to the stimulation of raw Pinellia ternata. After processing, the shape and contents of calcium oxalate changed, decreased respectively, and the stimulation of Pinellia ternata decreased. CONCLUSION: The needle crystal of calcium oxalate is one of the stimulating components in raw Pinellia ternata.