[Research on the origin and development of "Wu shan qi e"].

"Wu shan qi e"(five symptoms with good prognosis and seven with poor prognosis)is a theory used to determine the prognosis of external diseases in traditional Chinese medicine which have been mentioned in many ancient Chinese medicine books and have been valued by external doctors throughout history. However, it has been rarely discussed in modern literature. The theory were first seen in the Taiping Shenghui Fang, and the idea was originated from Ni Shun in Lingshu Yuban."Wu shan qi e"have evolved into many variants through the exertion of medical practitioners throughout history. By reviewing medical books of previous dynasties, it was found that there are two main versions: the Taiping Shenghui Fang version and its derivative versions, and the Waike Zhengzong version and its derivative versions.

[The definition of "Hu-Chang" in Chinese external medicine].

The Hu-Chang theory is widely used in Chinese external medicine diagnosis and treatment, and it has been documented in many archives and recordings. However, its provenance and connotation were still confusing and unclear. It was believed that The Covert Prescription for External Medicine (Mi Chuan Wai Ke Fang) by Zhao Yizhen in the late Yuan and early Ming Dynasty was the earliest work with the use of provenance from the existing records of Hu-Chang. In terms of its connotation, it was believed by many scholars that the part of speech of Hu-Chang was an adjective-noun phrase, which means "defensive barrier", ie, "local swelling area in surgical disease". However, based on the textual research and analysis of relevant ancient documents, Hu-Chang should be a verb-object phrase, which means "the vital Qi prevents the diseased area from being infected or deteriorated". In this sense, the characteristic of Hu-Chang, the local swelling range of surgical diseases, can be named as "supervising and monitoring the field barrier".

[A nested case-control study on the association of neonatal cord blood selenium, iron and copper with congenital heart disease].

Objective: To investigate the relationship between the levels of selenium, iron and copper in cord blood of neonates and the risk of congenital heart disease (CHD), and analyze their interaction effects. Methods: The subjects were obtained from the birth cohort in Lanzhou area established from 2010 to 2012. A baseline survey was conducted in the first trimester, and the follow-up was conducted in the second trimester, third trimester and 42 days after delivery. The umbilical vein blood was collected from newborns at delivery, and information on their birth outcomes was extracted from medical records. A nested case-control study was used to select 97 neonates with CHD newly diagnosed by echocardiography as the case group, and 194 neonates were selected as the control group by 1∶2 matching according to their mother's age, block and CHD onset time. Inductively coupled ion mass spectrometry was used to detect the concentrations of selenium, iron and copper in neonatal cord blood. The element exposure was categorized into three groups, the low, medium and high concentrations, according to the quartiles Q1 and Q3 of selenium, iron and copper concentrations in the control group. The association between cord blood selenium, iron and copper concentrations and CHD was analyzed by conditional logistic regression model using medium concentration as the reference standard. The association of their interactions with CHD was analyzed by a phase multiplication model. Results: The M (Q1, Q3) concentration of neonatal cord blood copper was 746.12 (467.48, 759.74) µg/L in the case group and 535.69 (425.21, 587.79) µg/L in the control group, with a statistically significant difference between the two groups (P<0.05). After adjustment for confounders, logistic regression models showed that the risk of CHD development was increased in neonates with either high copper in cord blood (OR=4.062, 95%CI: 2.013-8.199) or high copper combined with high iron (OR=3.226, 95%CI: 1.343-7.750). No correlation was observed between selenium and iron concentrations and the development of CHD in neonates. There was a multiplicative interaction between copper and iron in cord blood on the risk of developing CHD (OR=1.303, 95%CI: 1.056-1.608). Conclusion: There is a multiplicative interaction between iron and copper elements. The high copper and the high copper combined with high iron in umbilical cord blood are risk factors for neonatal CHD.

Low levels of organic compound trace elements improve the eggshell quality, antioxidant capacity, immune function, and mineral deposition of aged laying hens.

In the egg production industry, trace elements are required as additional dietary supplements to play vital roles in performance and egg quality. Compared to inorganic microelements (ITs), appropriate dose of organic trace microelements (OTs) are environmentally friendly and sufficient to satisfy the needs of hens. In order to evaluate the extent to which low-dose OTs replace whole ITs, the effects of organic copper, zinc, manganese, and iron compound on the performance, eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens were investigated. A total of 1 080 57-week-old Jing Hong laying hens were assigned to five groups with six replicates of 36 layers each for an 8-week experimental period. The birds were fed either a basal diet (control treatment (CT)) or the basal diet supplemented with commercial levels of inorganic trace elements (IT 100%) or the equivalent organic trace elements at 20%, 30%, and 50% of the inorganic elements (OT 20%, OT 30%, and OT 50%, respectively). Results showed that compared with those in the CT treatment, feeding hens with inorganic or organic microelement diet had significant effects on the eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens (P < 0.05). The eggshell strength and ratio between OT 30%, OT 50%, and IT 100% were similar at weeks 4 and 8, and the eggshell thickness of these groups was also similar at weeks 6 and 8. At week 8, the eggshell colour in OT 50% was darker than that in IT 100%. The mineral content in the eggshells of OT 50% and IT 100% significantly increased (P < 0.001), with no significant difference in effective thickness, mammillary thickness, and mammillary knob width between groups. There were no differences in the malondialdehyde content, total antioxidant capacity, and total superoxide dismutase activity in serum between OT 30%, OT 50%, and IT100%. While the catalase activities, the interleukin-1ß, interleukin-10, immunoglobulin G, and immunoglobulin M concentrations in serum were not significantly different between OT 50% and IT 100%. The mineral contents in the faeces of the organic groups were considerably reduced compared with those in IT 100% (P < 0.001). In conclusion, dietary supplementation with 30-50% organic compound microelements has the potential to replace 100% inorganic microelements in the hen industry for improving eggshell quality, mineral deposition in the eggshell, antioxidant capacity, and immune function, and reducing emissions to the environment without negative effects on laying performance.

Tributyrin supplementation in pasteurized waste milk: Effects on growth performance, health, and blood parameters of dairy calves.

This study evaluated the effects of incremental tributyrin supplementation in pasteurized waste milk on growth performance, health, and blood metabolism of dairy calves before and after weaning. Forty-eight newborn female Holstein dairy calves (39.6 ± 2.75 kg; mean ± standard deviation) were blocked by age and randomly assigned to 3 treatments: pasteurized waste milk (1) without supplementation, (2) with 1 g/L of tributyrin products (unprotected solid powder; containing 35% tributyrin), or (3) with 2 g/L of tributyrin products. The calves were weaned on d 56 and were raised until d 77. Data were analyzed for the preweaning, postweaning, and overall periods. The results showed that starter intake and hay intake were not different among treatments in any period of the trial, but the crude protein intake tended to increase linearly with tributyrin supplementation during the overall period. Although tributyrin supplementation had no effects on body weight during preweaning and overall periods, body weight increased linearly with tributyrin supplementation postweaning. The average daily gain tended to increase linearly during postweaning and overall periods. No effects were observed on feed efficiency in any period. A positive linear relationship between body length and tributyrin supplementation was observed during the postweaning period, but no differences were found for the other body structural measurements in any period. The results of diarrhea showed that tributyrin concentration had a negative linear relationship with diarrhea frequency during preweaning and overall periods. The rectal temperature did not differ among treatments in any period, but a treatment × week effect for rectal body temperature was observed. For blood metabolism, tributyrin supplementation had no effects on insulin, growth hormone, total protein, albumin, or globulin. No differences were found in serum amyloid A concentration in any of the periods, yet haptoglobin concentration decreased linearly with increasing tributyrin concentration during postweaning and overall periods. Endothelin concentration showed a tendency to decrease linearly during preweaning and postweaning periods and decreased linearly with tributyrin supplementation during the overall period. An increasing tributyrin concentration was associated with a negative linear relationship with IL-1ß concentration during the preweaning period, and no differences were found in the other periods. The concentration of IL-6 and tumor necrosis factor α were not different among treatments in any of the periods. These data suggest that increasing the concentration of tributyrin in pasteurized waste milk could increase growth performance and health of dairy calves, and incremental tributyrin supplementation could linearly reduce haptoglobin, endothelin, and IL-1ß concentrations, indicating a positive effect of tributyrin on alleviating oxidative stress and inflammatory status of dairy calves. Calves fed pasteurized waste milk supplemented with tributyrin products (containing 35% tributyrin) at 2 g/L compared with 1 g/L of milk had more improved growth and health.

Dietary supplementation of methionine mitigates oxidative stress in broilers under high stocking density.

We aimed to investigate whether dietary supplementation of methionine could mitigate intestinal oxidative injury in broilers under high stocking density (HSD). In the grower phase (d 22-42), 576 broilers with similar body weight were randomly chosen and divided into 8 groups in a 2 × 4 factorial experiment. Two different stocking densities (14 and 20 broilers per m2) were tested with 4 different methionine levels: 0.35%, 0.4%, 0.45%, or 0.5%. Intestinal morphological and oxidative stress markers were assessed at the end of the test period. The results showed that mortality of broilers was significantly higher in the HSD group fed 0.35% methionine diet than the other groups, which was reversed by supplementation with 0.40% to 0.50% methionine. HSD significantly decreased feed intake and daily weight gain. HSD treatment significantly decreased T-AOC, activity of GPX (P < 0.01) and increased the level of PCO (P < 0.01), MDA (P = 0.052) of plasma. The decreased glutathione peroxidase activity in the liver and jejunum caused by HSD was alleviated by additional methionine. Supplementation of methionine increased the ration of GSH/GSSG in the plasma. The jejunum villus height and ratio of villus height to crypt depth under low stocking density conditions with 0.40% methionine diet were the highest, whereas the 0.45% methionine group was the highest under HSD conditions. Thus, additional dietary supplementation of methionine mitigates oxidative stress in broilers under HSD conditions and 0.40% to 0.45% methionine can be applied in cage rearing broiler production for amelioration of oxidative stress caused by HSD.

[A probe into the origin and development of Ba Duan Jin's name and its connotation].

The earliest record of Ba Duan Jin is Yi Jian Zhi(, Records of Yi Jian) of the Song Dynasty, among which Ba Duan Jin refers to sitting Ba Duan Jin.Many Ba Duan Jin in books from Ming and Qing Dynasty, which are about nurturing vitality and preserving health, refer to the sitting Ba Duan Jin.Standing Ba Duan Jin was not called Ba Duan Jin at first. It was basically established in Dao Shu of the Song Dynasty, which has no specific name.At the procedure of spread, it was called "Yin Dao Jue" or "Lyu Zhen Ren An Le Ge" or "Xu Zhen Jun Yin Dao Jue" . Until the early years of Guangxu, when the Yijinjing Wai Jing Tu Shuo first call the standing Ba Duan Jin as Ba Duan Jin, the word Ba Duan Jin were mainly referred to standing Ba Duan Jin instead of sitting Ba Duan Jin.

Effects of the Zhikang capsule on healing of the flap after radical breast cancer surgery.

This study aimed to investigate the effect and mechanism of trauma flap healing promoted by the Zhikang capsule after radical breast cancer surgery. The enrolled breast cancer patients were randomly divided into two groups: treatment and observation. The patients in the treatment group were treated with the Zhikang capsule in addition to the conventional dressing changes, while patients in the observation group underwent only the regular dressing changes. Serum samples of 98 breast cancer patients (with complete clinical data) who underwent modified radical mastectomy were collected and analyzed for expressions of transforming growth factor beta (TGF-ß) and basic fibroblast growth factor (bFGF). The drainage fluid amount and tissue necrosis rate were found to be lower in the treatment group than in the observation group. Moreover, bFGF expression in peripheral blood was higher in the treatment group than in the observation group. However, no significant difference was found between the two groups in the expression of TGF-ß in peripheral blood. In conclusion, Zhikang capsule is effective in promoting flap healing after radical breast cancer surgery, and the increase of bFGF expression in peripheral blood may be the underlying mechanism.

Effect of curcumin on p38MAPK expression in DSS-induced murine ulcerative colitis.

The aim of this study was to determine the therapeutic effect of curcumin on dextran sulfate sodium-induced ulcerative colitis (UC) and to explore the related mechanism. Sixty mice were randomly divided into 6 groups. A group was the normal control group; B group was the model group; C group was the 1.5 mg/kg dexamethasone group based on the B group; and D, E and F groups were 15, 30, and 60 mg/kg curcumin groups, respectively, based on the B group. The mice were killed 7 days after treatment; the expression of TNF-α and MPO in colon tissue was determined with ELISA, and colon p-p38MAPK and p38MAPK mRNA expression was evaluated by immunohistochemistry and RT-PCR, respectively. In the C, D, E, and F groups, TNF-α and MPO levels significantly decreased (P < 0.05), and the expression of p-p38MAPK also significantly decreased (P < 0.01). The expression of p38MAPK mRNA in the C, D, E, and F groups decreased (P < 0.01), and there was a statistically significant difference between the E and F groups (P < 0.01). Curcumin had a therapeutic effect, which probably played a role in UC treatment by inhibiting the p38MAPK signaling pathway, thereby reducing the release of TNF-α.

Radix Astragali lowers kidney oxidative stress in diabetic rats treated with insulin.

Fluctuations in glucose levels in diabetic patients can result in oxidative stress, resulting in an increased risk for diabetic complications. We investigated whether antioxidation would protect the kidney from oxidative stress in diabetic rats treated with insulin and provide evidence for the efficacy of antioxidant treatment in diabetes management. Diabetes was induced by injection of Streptozotocin intraperitoneally in male Wistar rats. Diabetic rats received either insulin, both insulin and Radix Astragali (RA), RA, or no treatment. The levels of malondialdehyde (MDA), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) in kidney were determined. The changes of blood glucose levels and body weight were monitored. The levels of serum creatinine (Scr) were determined. The expression of PKCα was determined by western blot. NF-κB activation in kidney was assessed using EMSA. Compared to diabetic rats treated with insulin alone, the diabetic rats treated with combination of insulin and RA showed: (1) significantly lower levels of MDA, IL-6, TNF-α, and Scr (p < 0.05); (2) significantly higher SOD and GSH-Px activities (p < 0.05); (3) significantly lower NF-κB activation and lower expression levels of PKCα (p < 0.05); (4) significantly smaller kidney-to-body weight ratio (p < 0.05). RA is an effective agent in lowering oxidative stress in diabetic rats treated with insulin. Antioxidation is beneficial in reducing the risk of kidney damage due to oxidative stress in diabetic patients.

The effects of combined human parathyroid hormone (1-34) and zoledronic acid treatment on fracture healing in osteoporotic rats.

UNLABELLED: Ovariectomized (OVX) rats with tibial fracture received vehicle, ZA, PTH, or ZA plus PTH treatment for 4 and 8 weeks. Bone metabolism, callus formation, and the mass of undisturbed bone tissue were evaluated by serum analysis, histology, immunohistochemistry, radiography, micro-computerized tomography, and biomechanical test. INTRODUCTION: Previous studies have demonstrated the effect of ZA or PTH on osteoporotic fracture healing. However, reports about effects of ZA plus PTH on callus formation of osteoporotic fracture were limited. This study was designed to investigate the impact of combined treatment with ZA and PTH on fracture healing in OVX rats. METHODS: Twelve weeks after bilateral ovariectomy, all rats underwent unilateral transverse osteotomy on tibiae. Animals then randomly received vehicle, ZA (1.5 µg/kg weekly), PTH (60 µg/kg, three times a week), or ZA plus PTH until death at 4 and 8 weeks. The blood and bilateral tibiae of rats were harvested for evaluation. RESULTS: All treatments increased callus formation and strength other than the control; ZA + PTH showed the strongest effects on percent bone volume (BV/TV), trabecular thickness, total fluorescence-marked callus area, and biomechanical strength. Additionally, inhibited RANKL and enhanced osteoprotegerin expression were observed in the ZA + PTH group. But no difference in bone mineral density and BV/TV of the contralateral tibiae was observed between treated groups. CONCLUSION: Findings in this study suggested an additive effect of ZA and PTH on fracture healing in OVX rats, and this additive effect was specific to callus formation, not to undisturbed bone tissue.

Effect of several supplemental Chinese herbs additives on rumen fermentation, antioxidant function and nutrient digestibility in sheep.

Two experiments were carried out in this study. Experiment 1 was conducted to examine the effects of several supplemental Chinese herbs on antioxidant function and slaughtered body weight in sheep. Results indicated that Fructus Ligustri Lucidi supplementation improved the blood antioxidant function [higher concentration of glutathione reductase (GR), superoxide dismutase and lower concentration of malondialdehyde] and slaughtered body weight in sheep (p < 0.05). Experiment 2 was conducted to investigate the effect of Fructus Ligustri Lucidi extract (FLLE) on rumen fermentation and nutrient digestibility in sheep. Four levels of FLLE treatments, i.e. 0, 100, 300 and 500 mg/kg dry matter (DM), were used in this part. Addition of FLLE at 300 or 500 mg/kg DM increased total volatile fatty acid (VFA) concentration and propionate proportion, decreased ammonia-N concentration in the ruminal fluid, reduced blood urea nitrogen concentration at 2, 4, 6 and 8 h after morning feeding (p < 0.05). Addition of FLLE at all dosages had no effect on ruminal pH value and acetate concentration at all sampling time points in sheep (p > 0.05). Dynamic degradation coefficient c of maize DM was significantly increased by supplementing FLLE at 300 or 500 mg/kg DM (p < 0.05). Fructus Ligustri Lucidi extract addition had no effect on degradation coefficients a, b, c of DM and nitrogen of soybean meal; a, b of maize DM; a, b, c of maize nitrogen; and a, b, c of neutral detergent fibre (NDF) and acid detergent fibre (ADF) of Chinese wildrye (p > 0.05). Addition of FLLE at 300 or 500 mg/kg DM increased DM and organic matter digestibility of diet (p < 0.05). Fructus Ligustri Lucidi extract addition had no effect on digestibility of diet's NDF, ADF and crude protein (p > 0.05). From the aforementioned results, it is indicated that FLLE improved antioxidant status and slaughtered body weight. Fructus Ligustri Lucidi extract addition has capability to modulate rumen fermentation, increase the maize degradation rate, total volatile fatty acid concentration and propionate proportion in sheep.

Systemic treatment with strontium ranelate promotes tibial fracture healing in ovariectomized rats.

UNLABELLED: Systemic treatment with strontium ranelate (SR) was performed on ovariectomized (OVX) rats with fractured tibiae. Callus quality was assessed by radiographic, histological, micro-computerized tomography, and biomechanical examinations at 4 and 8 weeks after fracture. Results revealed that systemic applied SR promoted osteoporotic fracture healing. INTRODUCTION: Several studies have demonstrated the dual effect of SR on osteoporotic and undisturbed bone. However, reports of their effect on osteoporotic fracture healing are limited. This study was designed to investigate the effects of SR on bone regeneration in OVX rats with fractured tibiae. METHODS: Three months after being OVX, female Sprague-Dawley rats accepted bilateral osteotomy on proximal tibiae fixed with intramedullary wires and were divided into two groups: OVX and OVX + SR (625 mg/kg/day). Callus quality was evaluated at 4 and 8 weeks postfracture. RESULTS: Compared with OVX group, SR treatment significantly increased bone formation, BMD, biomechanical strength, and improved microstructural properties of the callus. The ultimate load was increased by 211.0% and 61.4% (p<0.01), and the total bone volume of callus by 74.8% and 79.3% (p<0.01) at 4 and 8 weeks postfracture, respectively. SR treatment also promoted healing progress with increased osteogenesis at 4 weeks; more mature and tightly arranged woven or lamellar bone at 8 weeks across the fracture gap in histological analysis. CONCLUSION: This study suggests that systemic treatment with strontium ranelate could promote tibial fracture healing in OVX rats.

Callus formation enhanced by BMP-7 ex vivo gene therapy during distraction osteogenesis in rats.

This study was designed to observe the effects of bone morphogenetic protein-7 (BMP-7) ex vivo gene therapy on callus formation during rat mandibular distraction osteogenesis (DO). Fifty-four Sprague-Dawley rats underwent osteodistraction of the right mandible and were then randomly divided into three groups. Immediately after distraction, autologous bone marrow mesenchymal stem cells (MSCs) transfected with BMP-7, MSCs untransfected with BMP-7, and physiological saline were injected into the distraction gaps of the mandibles in groups A, B, and C, respectively. Nine animals from each group were euthanized at 2 and 6 weeks after completion of distraction. The distracted mandibles were removed and processed for radiographic, histological, immunohistochemical, and scanning electron microscopic examinations as well as Ca/P ratio analysis. Group A animals showed greater bone formation and earlier mineralization in the distracted callus when compared with group B. Similarly increased callus formation was found in group B than group C. Positive immunostaining of BMP-7 was observed in the distracted callus in all groups. However, BMP-7 expression was much stronger in group A compared with groups B and C. The results of this study suggest that BMP-7-mediated ex vivo gene transfer based on MSCs may accelerate callus formation in distraction osteogenesis and facilitate consolidation. Local gene therapy may ultimately be an alternative or supplemental approach to DO enhancement, especially for patients whose osteogenic potentials are compromised by diseases such as osteoporosis, severe trauma, and postoncologic irradiation.

Sequestration of As by iron plaque on the roots of three rice (Oryza sativa L.) cultivars in a low-P soil with or without P fertilizer.

A pot experiment was carried out in a greenhouse to investigate the sequestration of As in iron plaques on root surface of three rice (Oryza sativa L.) cultivars. Phosphate (P) fertilization increased both plant biomass and tissue P concentrations significantly, indicating that the soils used in this study was highly P-deficient. Results from this study confirmed that low P supply improved the formation of iron plaque on rice roots. As a consequence, arsenic (As) concentrations in DCB-extracts with no P addition were significantly higher than those with P fertilization. Arsenic was highly sequestrated in iron plaque; arsenic concentration in iron was up to nearly 120 mg kg(-1), while arsenic concentrations in roots were just several mg kg(-1). Both arsenic and phosphate concentrations in iron plaque were highly positively correlated with the amounts of iron plaque (DCB-extractable Fe). Contrary to normal understanding that increasing P supply could reduced As accumulation in plants, results from the present study showed that P fertilization did not inhibit the As uptake by plants (As accumulation in aboveground), which was probably due to the fact that iron plaque formation was improved under low P conditions, thus leading to more As sequestration in the iron plaque. Thus results obtained in this study indicated that the iron plaque may inhibit the transfer of As from roots to shoots, and thus alter the P-As interaction in plant As uptake processes.

Adenoviral p53 gene therapy promotes heat-induced apoptosis in a nasopharyngeal carcinoma cell line.

BACKGROUND: It has previously been demonstrated that Ad5CMV-p53 gene transfer, either used alone or delivered concomitantly with ionizing radiation, resulted in cytotoxicity mediated by apoptosis in nasopharyngeal carcinoma (NPC) cell lines. In this study, a novel approach was evaluated of combining Ad5CMV-p53 gene therapy with hyperthermia (HT), in the CNE-1 NPC cell line, which harbours a mutation in codon 249 of the p53 gene. MATERIALS AND METHODS: CNE-1 cells were infected using either Ad5CMV-p53 or Ad5CMV-B-gal, followed, 24 h later, by HT (43 degrees C x 0-2 h). Protein was extracted for Western blot analysis, and apoptosis was evaluated using acridine-orange ethidium bromide staining, followed immediately by fluorescent microscopy examination for the proportion of cells displaying morphologic features of apoptosis. RESULTS: Ad5CMV-p53 gene therapy combined with HT resulted in a dose-dependent cytotoxicity with less than 1% clonogenic survival when 10 pfu/cell of Ad5CMV-p53 was combined with 2 h heating at 43 degrees C. Western blotting demonstrated that treatment with Ad5CMV-p53 resulted in the rapid expression of p53, which was minimally affected by HT. The inducible form of hsp70 was maximally expressed at 48 h post-HT, with minimal effect when cells were additionally treated with Ad5CMV-p53. Clonogenic cytotoxicity was associated with the development of apoptosis, with up to 70% of CNE-1 cells displaying morphologic features of apoptosis after the combination treatments. CONCLUSION: Based on the shapes of the clonogenic survival curves, Ad5CMV-p53 gene therapy and HT appear to interact in an additive manner, suggesting the therapeutic potential of this combined treatment approach for patients with NPC.

Folk medicine Terminalia catappa and its major tannin component, punicalagin, are effective against bleomycin-induced genotoxicity in Chinese hamster ovary cells.

Terminalia catappa L. is a popular folk medicine for preventing hepatoma and treating hepatitis in Taiwan. In this paper, we examined the protective effects of T. catappa leaf water extract (TCE) and its major tannin component, punicalagin, on bleomycin-induced genotoxicity in cultured Chinese hamster ovary cells. Pre-treatment with TCE or punicalagin prevented bleomycin-induced hgprt gene mutations and DNA strand breaks. TCE and punicalagin suppressed the generation of bleomycin-induced intracellular free radicals, identified as superoxides and hydrogen peroxides. The effectiveness of TCE and punicalagin against bleomycin-induced genotoxicity could be, at least in part, due to their antioxidative potentials.

Inhibition of protein kinase C activity promotes heat-induced apoptosis in RIF-1 but not in TR-4 cells.

PURPOSE: Inhibition of protein kinase C (PKC) activity has been demonstrated to reduce thermotolerance (TT), presumably by decreasing heat shock protein (HSP) production. Therefore, the interest was in evaluating this relationship further in two isogenic murine tumour cell lines: RIF-1 and its thermoresistant TR-4 selectant. MATERIALS AND METHODS: TT was induced in RIF-1 and TR-4 cells (45 degrees C for 15 min, then 37 degrees C for 6 h) with or without Ro31-8220, a specific inhibitor of PKC. PKC activity was assayed by determining the catalytic transfer of ATP to a specific substrate peptide. Survival was determined using the clonogenic assay. Apoptosis was quantitated by counting the number of cells demonstrating apoptosis after staining with acridine-orange/ ethidium bromide. Production of the inducible form of HSP70 was assessed using Western blot. RESULTS: At 2 microM Ro31-8220, >80% of PKC activity was inhibited in both cell lines, which was associated with no cytotoxicity at 37 degrees C. Basal HSP70 level was approximately 10-fold higher in the TR-4 compared with the RIF-1 cells. Upon TT induction, HSP70 level increased significantly in both cell lines, which was suppressed in the presence of Ro31-8220, but the relative amount of HSP70 remained high in the TR-4 cells. At 24 h, heat-induced apoptosis increased from 4 to 38% in RIF-1 cells in the presence of Ro31-8220, which was associated with a 26% reduction in clonogenic survival after thermotolerant heating. In contrast, <1% of TR-4 cells demonstrated apoptosis even with the highest dose of Ro31-8220, and no effect on survival was observed. CONCLUSION: Inhibition of PKC activity reduces HSP70 induction, which in turn is associated with promotion of heat-induced apoptosis in RIF-1 cells. However, the survival signals in the TR-4 cells are so strong, that even 80% inhibition of PKC activity has minimal impact on heat-induced apoptosis and survival in this thermoresistant cell line.