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This study explored toxicity attenuation processing technology of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction for the first time, and further explored its detoxification mechanism. Nine processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction were prepared by orthogonal experiment with three factors and three levels. Based on the decrease in the content of the main hepatotoxic component diosbulbin B before and after processing of Rhizoma Dioscoreae Bulbiferae by high-performance liquid chromatography, the toxicity attenuation technology was preliminarily screened out. On this basis, the raw and representative processed products of Rhizoma Dioscoreae Bulbiferae were given to mice by gavage with 2 g·kg~(-1)(equival to clinical equivalent dose) for 21 d. The serum and liver tissues were collected after the last administration for 24 h. The serum biochemical indexes reflecting liver function and liver histopathology were combined to further screen out and verify the proces-sing technology. Then, the lipid peroxidation and antioxidant indexes of liver tissue were detected by kit method, and the expressions of NADPH quinone oxidoreductase 1(NQO1) and glutamate-cysteine ligase(GCLM) in mice liver were detected by Western blot to further explore detoxification mechanism. The results showed that the processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction reduced the content of diosbulbin B and improved the liver injury induced by Rhizoma Dioscoreae Bul-biferae to varying degrees, and the processing technology of A_2B_2C_3 reduced the excessive levels of alanine transaminase(ALT) and aspartate transaminase(AST) induced by raw Rhizoma Dioscoreae Bulbiferae by 50.2% and 42.4%, respectively(P<0.01, P<0.01). The processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction reversed the decrease protein expression levels of NQO1 and GCLM in the liver of mice induced by raw Rhizoma Dioscoreae Bulbiferae to varying degrees(P<0.05 or P<0.01), and it also reversed the increasing level of malondialdehyde(MDA) and the decreasing levels of glutathione(GSH), glutathione peroxidase(GPX), and glutathione S-transferase(GST) in the liver of mice(P<0.05 or P<0.01). In summary, this study shows that the optimal toxicity attenuation processing technology of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction is A_2B_2C_3, that is, 10% of Paeoniae Radix Alba decoction is used for moistening Rhizoma Dioscoreae Bulbiferae and processed at 130 ℃ for 11 min. The detoxification mechanism involves enhancing the expression levels of NQO1 and GCLM antio-xidant proteins and related antioxidant enzymes in the liver.
Subject(s)
Mice , Animals , Antioxidants/analysis , Plant Extracts/pharmacology , Drugs, Chinese Herbal/chemistry , Rhizome/chemistry , Paeonia/chemistry , Glutathione/analysisABSTRACT
On the basis of the previous work of the research group, the orthogonal design method was further used to optimize the processing technology for reducing toxicity of fried Tripterygium wilfordii in Lysimachia christinae Decoction. A total of 9 processed products of T.wilfordii in L.christinae decoction were prepared by four factors and three levels orthogonal design table. The contents of triptolide in T.wilfordii were determined by high performance liquid chromatography(HPLC) before and after processing: 4.27, 3.92, 3.57, 2.75, 2.42, 2.66, 3.51, 1.87, 1.75, 2.03 μg·g~(-1). On this basis, the above processed products were orally given to mice for 28 days. 12 hours after the last administration, food fasting except water was provided, and 24 hours later, the eyeballs were taken for blood and liver tissue. Serum biochemical indexes, liver lipid peroxidation and antioxidant related indexes were detected by kit method. Twenty-eight days after oral administration of raw T.wilfordii, the levels of serum alanine aminotransferase(AST), alanine aminotransferase(ALT), alkaline phosphatase(ALP) and liver malondialdehyde(MDA) in mice increased by 91%(P<0.01), 46%(P<0.05), 73%(P<0.01) and 99%(P<0.01), while the liver antioxidant indexes such as superoxide dismutase(SOD), glutathione(GSH), glutathione peroxidase(GPX) and glutathione-S transferase(GST) significantly decreased(P<0.01). After administration of the processed products, the above indexes were significantly reversed(P<0.01 or P<0.05). Especially, the processing conditions of A_3B_2C_1D_3 had the best detoxification effect on T.wilfordii, which decreased the high levels of AST, ALT, ALP and MDA by 49%(P<0.01), 32%(P<0.01), 42%(P<0.01), and 17%(P<0.05). Therefore, the best processing conditions for T.wilfordii in L.christinae decoction were A_3B_2C_1D_3, namely "15% mass fraction of L.christinae, 1 h moistening time, 160 ℃ frying temperature, and 9 min frying time".
Subject(s)
Animals , Mice , Antioxidants , Liver , Primulaceae , Technology , TripterygiumABSTRACT
Objective:To investigate the effect of different extracts of Lysimachiae Herba on the main toxicity induced by Tripterygii Radix et Rhizoma. Method:Ninety male SPF Kunming mice were randomly divided into 9 groups according to their body weight，control group， Lysimachiae Herba water extract group， Lysimachiae Herba 30% ethanol extract group， Tripterygii Radix et Rhizoma group， Tripterygii Radix et Rhizoma combined with Lysimachiae Herba water extract group， Tripterygii Radix et Rhizoma combined with Lysimachiae Herba 30% ethanol extract group， Tripterygii Radix et Rhizoma combined with Lysimachiae Herba 60% ethanol extract group， Tripterygii Radix et Rhizoma combined with Lysimachiae Herba 95% ethanol extract group and Tripterygii Radix et Rhizoma combined with Lysimachiae Herba ethyl acetate extract group. The dosage of Tripterygii Radix et Rhizoma and Lysimachiae Herba were 2，1 g·kg<sup>-1</sup> based on crude drugs， respectively. The control group was given an equal volume of solvent， and each group was given by gavage for 14 consecutive days. The blood and liver tissues were taken 24 hours after the last administration. The enzyme linked immunosorbent assay （ELISA） was used to detect serum biochemical indexes and liver lipid peroxidation/antioxidant indexes in mice. Meanwhile， principal component analysis was used to evaluate the attenuating effect and the mechanism of Lysimachiae Herba extract on toxicity of Tripterygii Radix et Rhizoma. Result:Compared with control group， Tripterygii Radix et Rhizoma caused the levels of alanine aminotransferase（ALT），aspartic acid amino transferase（AST），alkaline phosphatase（ALP） in serum of mice， and the levels of malondialdehyde （MDA） in liver， and comprehensive score of toxicity （Z value） produced by the above four indexes increased significantly （<italic>P</italic><0.01）. The levels of total superoxide dismutase （T-SOD），glutathione-peroxidase （GPX），glutathione-S transferase （GST） decreased significantly （<italic>P</italic><0.01） in liver. Compared with Tripterygii Radix et Rhizoma group， after intervention with extracts of two solvents （water， 30% ethanol） of Lysimachiae Herba， the levels of serum ALT， AST， ALP and liver MDA were significantly decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， while the levels of liver T-SOD， GPX and GST were significantly increased （<italic>P</italic><0.01）. After intervention with extracts of two solvents （60% ethanol， 95% ethanol） of Lysimachiae Herba， the levels of serum ALT， AST， ALP were significantly decreased （<italic>P</italic><0.01）， and liver GPX levels were significantly increased （<italic>P</italic><0.01）. After the intervention with ethyl acetate extract of Lysimachiae Herba， only the level of serum AST was significantly decreased （<italic>P</italic><0.05） and the level of GPX was significantly increased （<italic>P</italic><0.05）. After the intervention with extracts of different solvents （water， 30% ethanol， 60% ethanol， 95% ethanol， ethyl acetate） of Lysimachiae Herba， it can significantly reduce the comprehensive score of toxicity （<italic>P</italic><0.01）. The overall decline rates of toxicity were 127.5%， 113.4%， 98.1%， 56.3% and 31.0% respectively. Among them， the toxicity reduction rate of the extracts with water as a solvent was 14.1%， 29.4%， 71.2%， 96.5% higher than those of other solvent extracts with ethanol. Conclusion:The extracts of different solvents （water， 30% ethanol， 60% ethanol， 95% ethanol and ethyl acetate） of Lysimachiae Herba can reverse the toxicity induced by Tripterygii Radix et Rhizoma in varying degrees. Among them， water and 30% ethanol are the best solvents for detoxification， especially water as the extraction solvent， and with the increase of ethanol content or fat solubility of extraction solvent， the detoxification shows a downward trend.
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OBJECTIVE: To investigate the effects of different processing methods on antitussive and hepatotoxicity of Dioscorea bulbifera L.. METHODS The raw and processed products of Dioscorea bulbifera L., fried Dioscorea bulbifera L. (FD), wine-fried Dioscorea bulbifera L. (WD), liquorice liquid-fried Dioscorea bulbifera L. (LD) and angelicae liquid-fried Dioscorea bulbifera L. (AD) were administered to mice at a dose of 1.7 g•kg-1 for 7 d. The effects of processing on the cough relief of Dioscorea bulbifera L. were evaluated by the cough test induced by concentrated ammonia water. The detoxification effect and the preliminary mechanism of processing on Dioscorea bulbifera L. were evaluated by the detection and analysis of the relevant indicators. RESULTS: Compared with the RD group, LD and AD group had longer cough latency (P0.05). Compared with the AD group, there were more coughing times in LD group (P0.05) in mice induced by concentrated ammonia water. The intervention of WD only had significant reversal effect on the low GST level of liver (P0.05). CONCLUSION: Processing with Angelica sinensis and licorice could enhance the antitussive effect of Dioscorea bulbifera L. and reduce its hepatotoxicity. The mechanism of attenuation may be related to the inhibition of lipid peroxidation and the enhancement of antioxidant level in liver. The antitussive effect of two kinds of processing, stir-fried and wine-fried, is only effective, but no synergistic effect, no toxic effect and no attenuation effect are found.
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To analyze the reasons of impaired vision after LASlK and explore its preventive treatment measures preliminarily.METHODS: ln this retrospective study, 175 eyes of 134 patients whose vision was decreased after LASlK were included. The constituent ratio of every reason was counted and uncorrected visual acuity ( UCVA ) between pre-treatment and post-treatment were compared by paired t-test respectively.RESULTS:The overall incidence of impaired vision after LASlK was 1. 86%. The constituent ratio of regression was 51. 43% and UCVA increased from 0. 61±0. 22 to 0. 90±0. 38 (t=8. 00, P<0. 001) after treatment. The constituent ratio of punctate corneal epithelial defect was 32. 57% and UCVA increased from 0. 60±0. 19 to 1. 20±0. 24 (t=20. 00, P<0. 001 ) after treatment. The constituent ratio of accommodative spasm was 5. 14% and UCVA increased from 0.76±0. 21 to 1. 32±0. 22 (t=8. 14, P<0. 001) after treatment. The constituent ratio of corneal flap shift and gauffer was 4% and UCVA increased from 0. 29 ± 0. 26 to 1. 24 ± 0. 28 ( t = 6. 33, P<0. 001 ) after treatment. The constituent ratio of corticosteroid - induced ocular hypertension was 4% and UCVA increased from 0. 57±0. 05 to 1. 0 ± 0. 16 ( t= 2. 53, P<0. 05 ) after treatment. The constituent ratio of fundus lesions and diffuse lamellar keratitis ( DLK) was 2. 86% and UCVA all increased by different degrees after treatment.CONCLUSlON: The reasons of impaired vision after LASlK are many and varied. These cases could recover their vision by discovery and treatment in time, and the appropriate preventive measures were essential.
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BACKGROUND: The quality of treatment for critically ill children varies widely at different hospitals. This study aimed to analyze the characteristics of mortality in a pediatric emergency department (PED) at a tertiary children's hospital in Guangzhou, China and to investigate the risk factors associated with the mortality. METHODS: The mortality of pediatric patients at the hospital from 2011 to 2013 was retrospectively analyzed using descriptive statistics. RESULTS: Altogether 466919 patients visited the PED during the period and 43925 of them were admitted for further observation. In 230 deaths, the ratio of boys to girls was 1.4:1, and their age ranged from 2 hours to 16 years (median, 5 months). The time from admission to death ranged from 0 to 216 hours (median, 1.5 hours). There were 92 (40％) patients who died within 24 hours after admission and 104 (45.2％) patients who died on arrival. The prominent causes of the deaths were respiratory diseases, neuromuscular disorders, cardiovascular diseases, and sepsis, most of which were ascribed to severe infection. Sixty-five deaths were associated with more than one concomitant problem. The top concomitant problems were congenital malformation, low gestational age, and severe birth asphyxia. CONCLUSIONS: In our center, 40％ of the patients in the PED died of fatal acute diseases, and pneumonia was the first leading cause of the deaths. Almost half of the deaths occurred on arrival and the rest were due to end-stage malignant diseases. This study emphasized the importance of prevention of birth deficits by reducing deaths in infants and children.
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<p><b>OBJECTIVE</b>To evaluate electroacupuncture on the efficacy and the safety of gastric and systemic functional recovery in patients with carbon dioxide pneumoperitoneum undergoing gynaecological laparoscopic surgery.</p><p><b>METHODS</b>Sixty patients undergoing gynaecological laparoscopic surgery were randomly divided into a regular anesthesia group (group A) and a combination electroacupuncture anesthesia group (group B), 30 cases in each group. The group A were treated with regular general anesthesia, and on the basis of general anesthesia, the group B were treated with electroacupuncture at bilateral Zusanli (ST 36) and Liangqiu (ST 34) by continuous wave and a frequency of 2 Hz 30 minutes before the surgery until the end of the operation. Sedation scale by Ramsay, nausea, vomiting and mental state scores of postoperative were evaluated and compared between the two groups.</p><p><b>RESULTS</b>Compared with the group A, the sedation scale (1.19 +/- 0.56 vs 1.96 +/- 0.90), nausea score (0.59 +/- 0.80 vs 1.81 +/- 0.88), vomiting score (0.48 +/- 0.94 vs 1.89 +/- 0.93) and mental state score (0.96 +/- 0.73 vs 1.69 +/- 0.75) were all decreased significantly in the group B (all P < 0.01).</p><p><b>CONCLUSION</b>Combination electroacupuncture anesthesia can relieve postoperative irritable, nausea and vomiting, improve mental state after the operation and promote the gastric and systemic functional recovery. It is a good method for the adjustment of carbon dioxide pneumoperitoneum during laparoscopic surgery.</p>
Subject(s)
Adult , Female , Humans , Middle Aged , Young Adult , Electroacupuncture , Genital Diseases, Female , General Surgery , Gynecologic Surgical Procedures , Laparoscopy , Postoperative Complications , Therapeutics , Postoperative Period , Recovery of FunctionABSTRACT
<p><b>OBJECTIVE</b>To investigate the feasibility of Matrix-Assisted Laser Desorption-Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) , according to the genetic test of non-syndromic hearing loss (NSHL), and check using the direct sequencing.</p><p><b>METHODS</b>Peripheral blood was collected from 454 NSHL patients. DNA samples were extracted and 20 loci of the four common disease-causing genes were analysed by MALDI-TOF-MS, including GJB2 (35delG, 167delT, 176_191del16, 235delC, 299_300delAT ), GJB3 (538C→T, 547G→A), SLC26A4 (281C→T, 589G→A, IVS7-2A→G, 1174A→T, 1226G→A, 1229C→T, IVS15+5G→A, 1975G→C, 2027T→A, 2162C→T, 2168A→G), and mitochondrial 12S rRNA (1494C→T, 1555A→G). Direct sequencing was also used to analyse the aforementioned 20 loci in order to validate the accuracy of MALDI-TOF-MS.</p><p><b>RESULTS</b>Among the 454 patients, 166 cases (36.56%) of disease-causing mutations were detected, which included 69 cases (21.15%) of GJB2 gene mutation, four cases (0.88%) of GJB3 gene mutation, 64 cases (14.10%) of SLC26A4 gene mutation, and three cases (0.66%) of mitochondrial 12S rRNA gene mutation. Moreover, the results obtained from direct sequencing and MALDI-TOF-MS were consistent, and the results showed that the two methods were consistent.</p><p><b>CONCLUSIONS</b>The MALDI-TOF-MS detection method was designed based on the hearing loss-related mutation hotspots seen in the Chinese population, and it has a high detection rate for NSHL related mutations. In comparison to the conventional detection methods, MALDI-TOF-MS has the following advantages: more detection sites, greater coverage, accurate, high throughput and low cost. Therefore, this method is capable of satisfying the needs of clinical detection for hearing impairment and it is suitable for large-scale implementation.</p>
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Young Adult , Asian People , Genetics , Connexins , Deafness , Genetics , Genetic Testing , Mutation , Oligonucleotide Array Sequence Analysis , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of RKIP, p65 and pERK in hepatocellular carcinoma (HCC) and theIr correlation with invasion and metastasis of HCC.</p><p><b>METHODS</b>Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of RKIP mRNA. The expression levels of RKIP, p65 and pERK proteins in HCC tumor and peritumoral tissues were determined by immunohistochemistry and Western blot analysis. Statistical analysis was performed to determine the relationship between their expression and clinicopathological parameters.</p><p><b>RESULTS</b>RKIP protein expression level (RKIP/actin) was 0.579 ± 0.380 in HCCs, 1.178 ± 0.659 in peritumoral tissues and 1.115 ± 0.442 in normal liver tissues. The pERK protein level was 1.023 ± 0.478, 0.605 ± 0.367 and 0.461 ± 0.293, p65 protein level was 0.83 ± 0.376, 0.63 ± 0.337 and 0.466 ± 0.345, respectively. Immunohistochemistry analysis showed that the RKIP positive rates in HCCs, peritumoral tissues and normal liver tissues, were 22.2%, 86.0%, and 93.8%, positive rates of p65 were 73.6%, 56.0% and 37.5%, positive rates of pERK were 65.3%, 38.0% and 31.3%, respectively. Statistical analysis revealed that there was a significant difference in RKIP protein expression levels (P < 0.05), but no significant difference in RKIP mRNA expression levels (P > 0.05) among HCC tumors, peritumoral tissues and normal liver tissues. The p65-positive and pERK-positive rates were higher in tumor tissues than that in peritumoral tissues and in normal liver tissues (P < 0.05), but RKIP-positive rates were lower in tumor tissues than that in paritumoral tissues and normal liver tissues (P < 0.05). RKIP protein expression levels were significantly lower in HCCs with intrahepatic or lymphatic metastasis than that in without. The RKIP positive rates in moderately and well differentiated HCCs were significantly higher than that in poorly differentiated HCCs. There was a relationship between RKIP and pERK expressions (P = 0.04), but RKIP expression was not correlated with p65 expression in HCCs (P = 0.143).</p><p><b>CONCLUSIONS</b>Our findings indicate that the down-regulation of RKIP expression may serve as a predictive marker for HCC development, progression and metastasis, which may contribute to the elevated ERK activity. The inhibiting effect of RKIP on invasion and metastasis of liver cancer cells may be due to the down-regulation of pERK expression rather than p65 expression.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Metabolism , Pathology , Down-Regulation , Extracellular Signal-Regulated MAP Kinases , Metabolism , Gene Expression Regulation, Neoplastic , Liver , Metabolism , Liver Neoplasms , Metabolism , Pathology , Lymphatic Metastasis , Neoplasm Invasiveness , Phosphatidylethanolamine Binding Protein , Genetics , Metabolism , Phosphorylation , RNA, Messenger , Metabolism , Transcription Factor RelA , MetabolismABSTRACT
Objective: To investigate the expression and clinical significance of Raf kinase inhibitor protein (RKIP) and phosphorylated-extracellular signal regulated kinase (P-ERK) in hepatocellular carcinoma (HCC) and their correlations with the invasion and metastasis of HCC. Methods: Immunohistochemistry was used to detect the expressions of RKIP and P-ERK in HCC tissues, paracancerous tissues, and normal liver tissues. Statistical analysis was used to determine the relationship between their expressions and clinicopathological parameters. Results: Statistical analysis revealed that RKIP-positive staining rate was lower in tumor tissues than that in tumor-surrounding tissues and in normal liver tissues. But P-ERK-positive staining rate was significantly higher in tumor tissues than that in tumor-surrounding tissues and normal liver tissues. The difference was significant. RKIP expression had negative correlation with P-ERK expression (r = - 0.227, P = 0.039); RKIP expression in HCC was associated with intrahepatic or lymphatic metastasis and tumor differentiation (P < 0.05). P-ERK expression in HCC was related with tumor differentiation, tumor thrombosis, and intrahepatic or lymphatic metastasis (P < 0.05). Conclusions: These findings indicate that down-regulation or loss of RKIP expression is closely related with HCC development and progression. The down-regulation or loss of RKIP expression stimulates migration and metastasis of HCC by up-regulating P-ERK expression.
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0.05), and revealed highly significant difference compared with those of water samples collected from families of GI group with 1_year residential history (P