Objective To investigate the expression of Notch4 protein and to analyze its correlation with the clinical parameters and the microvessel dentisty in renal cell carcinoma. Methods The expression of Notch4 was examined in 60 cases of renal cell carcinoma and the para-carcinoma tissue by SP immunohistochemical stain-ing ,and CD34 detection was used for counting microvessel density. Statistical analysis was performed to reveal the correlation with clinicopathological parameters ,microvessel density and prognosis. Results The positive rate of Notch4 protein expression was 75%(45/60)in para-carcinoma tissue,and was 43.3%(26/60)in renal cell car-cinoma,with significant difference on tumor grade and Lymph node metastasis(P<0.05). The microvessel densi-ty in Notch4 positive tissues was significant lower than that in the negative samples(P<0.05). The survival time of patients with Notch4 positive expression was significantly longer than that of patients with Notch4 negative expres-sion(P<0.05). Conclusion Notch4 protein plays an important role in the development of renal cell carcinoma. Notch4 expression might both attenuate the malignant biological characteristics and suppress the angiogenesis dur-ing tumor development.

Objective: Recently hepatotoxicity induced by Polygoni Multiflori Radix Praeparata (PMRP) frequently happened. To aim at dealing with the problems of latest increase of online shopping PMRP and lack of quality monitoring mechanism, the quality and hepatotoxicity of various batches of online shopping PMRP were analyzed, the toxicity-attenuation effects of various processing technologies of Polygoni Multiflori Radix (PMR) were compared, and the reference for clinical safe medication of PMR was provided. Methods: The quality variance of these PMRP was evaluated and the hepatotoxicity-attenuation effects of various processing technologies were compared by adopting index component content determination, chemical fingerprint, and liver cell toxicity evaluation. Results: The content of diphenylethylene glycosides in 26 batches of online shopping PMRP was 0.004%-3.442%, therein eight batches could not conform to Chinese Pharmacopoeia 2010; The chemical fingerprint similarity was between 0.052 and 0.998, therein great differences among six batches existed; The hepatotoxicity difference was significant as 9.7 times, therein the toxicity of four groups of online shopping PMRP samples was higher than that of PMR medicinal material. The toxicity of PMRP attenuated slowly using atmospheric pressure steaming method, and the toxicity by steaming 12 h declined only 13.6%; The toxicity of PMRP attenuated relatively fast by high-pressure steaming and high-pressure black soya bean steaming methods, and the toxicity by steaming 5-6 h declined by 22.1% and tended towards stability. Conclusion: The quality of online shopping PMRP varies greatly and inadequate processing method increases the liver poisoning risk of PMRP to some extent, which indicates that the researches on technology standardization of PMRP should be strengthened. As well, the high-pressure steaming method can ensure the safety of PMRP and shorten the processing time greatly, which should be expected as a promising processing method.

BACKGROUND:Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE:To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS:The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×105 293T cells per wel , total 500μL. In the control group, 500μL culture medium was added;in the other three groups, 500μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION:By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins.

To investigate the changes of endocrine, cyclic nucleotide and immune systems in subjects of yang deficiency constitution, and to explore the relationship among characteristics and causes of yang deficiency constitution, the physiological and biochemical parameters.

The dosage-efficacy/toxicity relationship of the 50% alcohol extracts of Polygonum multiflorum was comparatively investigated on either normal or CCl4-induced chronic liver injury rats, by determining the general condition, serum biochemical indices and liver histopathology, coupled with the factor analysis. The dosages were 10 and 20 g raw materials per kg body weight. Compared with the normal control group, the normal high dose group showed significant increases of the serum alanine transaminase (ALT), total bilirubin (TBIL), high mobility group box 1 (HMGB-1) and interleukin-1β (IL-1β) (P < 0.05 or P < 0.01), as well the frequent incidences of inflammatory cell infiltration, hepatic sinus enlargement and fiber stripes formation in histopathological sections. Compared with the model control group, the model low dose group showed significant declines of serum ALT, aspartate transaminase (AST) and total bile acid (TBA) (P < 0.05), as well the alleviation of vacuoles of hepatocytes, but no amelioration of the inflammatory cell infiltration and fibrous tissue hyperplasia; moreover, the model high dose group showed significant degeneration declines of serum HMGB-1, tumor necrosis factor-α (TNF-α) and IL-1β (P < 0.05, P < 0.01), as well the evident alleviation of vacuoles degeneration of hepatocytes, inflammatory cells infiltration and fibrosis degree. The factor analysis showed that the low dosage treatment had almost neither injuring effect on the normal rats nor protective effect on the model rats; while the high dosage treatment showed observable injuring effect on the normal rats, expressed by the significant increases of the factor-1 (HMGB-1, TNF-α and IL-1β as the main contributors) and factor-2 (TBIL, ALT and TBA as the main contributors) relative to the normal control group. The liver protective effect of the high dosage treatment could be observed with the significant reduction of the factor-1, indicating the effective alleviation of the expression of inflammatory cytokines. In conclusion, it could illustrated the phenomenon of symptom-based prescription theory of Polygonum multiflorum on rat livers: the high dosage of the herb had either an injuring effect on normal rats, or a therapeutic effect on the rats with chronic liver injury.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Bile Acids and Salts ; metabolism ; Bilirubin ; blood ; Chemical and Drug Induced Liver Injury ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Fallopia multiflora ; chemistry ; HMGB1 Protein ; metabolism ; Hepatocytes ; drug effects ; Interleukin-1beta ; metabolism ; Liver ; drug effects ; pathology ; Plant Extracts ; pharmacology ; Rats ; Tumor Necrosis Factor-alpha ; metabolism

Objective:To explore the relationship between somatic symptoms of major depressive disorder(MDD)and cortisol(COR) rhythm, C-reactive protein(CRP) and other immune-metabolism-related indicators, and understand its mechanism from the perspective of endocrine and immune regulation.Methods:A case-control study was conducted in hospitalized patients with MDD who met DSM-5 diagnostic criteria.According to the Patient Health Questionnaire (PHQ-15), PHQ-15 ≥10 were classified as the somatic major depressive disorder group(S-MDD group) and 73 patients were enrolled.PHQ-15 <5 was classified as the non-somatic depressive disorder group (NS-MDD group) and 70 patients were enrolled.Plasma cortisol (COR8, COR16 and COR24) levels were measured at 8∶00, 16∶00 and 24∶00 on the same day, plasma CRP and interleukin-6 (IL-6) level, serum uric acid (UA), blood glucose (GLU), blood lipid (TC, TG, HDL, LDL) level were detected at 8∶00.Independent sample t test, non-parametric test, chi-square test, repeated ANOVA, covariance analysis, and multivariate Logistic regression were used for statistical analysis. Results:①Time effect, grouping effect and the interaction effect of the time and grouping in the level of COR were statistically significant ( P<0.05). Covariance analysis excluded age as an influential factor, COR16, AUC(total cortisol output/area under the curve, AUC) and COR8-16 in S-MDD group ((90.50±40.57)μg/L, (1 425.12±564.78), (-6.43±5.76))were higher than those in NS-MDD group((68.74±31.51)μg/L, (1 251.57±456.61), (-8.77±5.48)), and the difference was statistically significant ( F=8.971, 4.320, 8.731, P<0.05). ②CRP in S-MDD group ((1.41±1.06)mg/L) were higher than that in NS-MDD group((0.61±0.53)mg/L), and the difference was statistically significant ( F=25.436, P<0.05). The proportion of patients with higher CRP level(CRP≥1 mg/L) in S-MDD group(58%) was higher than that in NS-MDD group(23%), and the difference was statistically significant(χ 2=17.824, P<0.01). ③Multivariate logistic regression analysis found that CRP ( OR=4.953, 95% CI: 2.407-10.193), COR8-16 ( OR=3.451, 95% CI: 1.380-8.633) were main risk factors of somatic symptoms of MDD ( P<0.05). Conclusion:Cortisol rhythm disturbance and high CRP level may be the biological basis of somatic symptoms in patients with MDD.

To investigate the difference of liver injury in rats gavaged with crude and processed Polygoni Multiflori Radix. The 75% ethanol extract of crude and processed Polygoni Multiflori Radix (50 g · kg(-1) crude medicine weight/body weight) were continuous oral administered to rats for 6 weeks. Serum biochemical indicators were dynamically detected, the change of liver histopathology was assessed 6 weeks later. Principal component analysis (PCA) was adopted to screen sensitive indicator of the liver damage induced by polygoni multiflori radix. Biochemical tests showed that the crude Polygoni Multiflori Radix group had significant increase of serum ALT, AST, ALP, DBIL and TBIL (P < 0.01 or P < 0.05) and significant decreases of serum IBIL and TBA (P < 0.01 or P < 0.05), while the processed Polygoni Multiflori Radix group showed no obvious changes, compared to the untreated normal group. Histopathologic analysis revealed that crude Polygoni Multiflori Radix group exhibited significant inflammatory cells infiltration in portal area around the blood vessels, tissue destruction and local necrosis of liver cells. There were not obvious pathological changes in processed Polygoni Multiflori Radix group. The results demonstrated that the injury effect of processed Polygoni Multiflori Radix on liver injury of rats was significantly lower than that of unprocessed, and that processing can effectively reduce the hepatotoxicity of Polygoni Multiflori Radix. Traditional transaminase liver function indicators were not sensitive for crude Polygoni Multiflori Radix induced liver damage. The serum content of DBIL and TBIL can reflect the liver damage induced by crude Polygoni Multiflori Radix early and can be sensitive indicators for clinical monitoring the usage of it.
Animals ; Chemical and Drug Induced Liver Injury ; etiology ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Female ; Liver ; drug effects ; injuries ; Male ; Plant Roots ; chemistry ; toxicity ; Polygonum ; chemistry ; toxicity ; Rats

The liver injury induced by Polygonum multiflorum Thunb. (PM) was investigated based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose. Sprague-Dawley (SD) rats were intragastrically administered with three doses (18.9, 37.8, 75.6 g crude drug per kg body weight) of 50% alcohol extracts of PM alone or co-treated with non-toxic dose of LPS (2.8 mg·kg(-1)) via tail vein injection. The plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were assayed and the isolated livers were evaluated for histopathological changes. The dose-toxicity relationships of single treatment of PM or co-treatment of LPS were investigated comparatively to elucidate the idiosyncratic hepatotoxicity of PM. The results showed that no significant alterations of plasma ALT and AST activities were observed in the groups of solo-administration of LPS (2.8 mg·kg(-1), i.v.) or different dosage (18.9, 37.8 and 75.6 g·kg(-1), i.g.) of PM, compared to normal control group (P > 0.05); while significant elevations were observed in the co-administration groups of PM and LPS. Treatment with LPS alone caused slight infiltration of inflammatory cells in portal area but no evident hepatocytes injury. Co-treatment with LPS and PM (75.6 g·kg(-1), i.g.) caused hepatocyte focal necrosis, loss of central vein intima and a large number of inflammatory cell infiltration in portal areas. When further reduce the dosage of PM, significant increases of plasma ALT and AST activities (P < 0.05) were still observed in co-administration groups of LPS and PM (1.08 or 2.16 g·kg(-1)), but not in LPS or PM solo-administration groups. Nevertheless, the co-treatment of low dosage of PM (0.54 g·kg(-1)) with LPS did not induce any alteration of plasma ALT and AST. In conclusion, intragastric administration with 75.6 g·kg(-1) of PM did not induce liver injury in normal rats model; while the 2 folds of clinical equivalent dose of PM (1.08 g·kg(-1)) could result in liver injury in the LPS-based idiosyncratic hepatotoxicity model, which could be used to evaluate the idiosyncratic hepatotoxicity of PM.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Chemical and Drug Induced Liver Injury ; pathology ; Hepatocytes ; pathology ; Lipopolysaccharides ; Polygonum ; toxicity ; Rats ; Rats, Sprague-Dawley

UNLABELLED: The DNA barcoding method was used to accurately and rapidly identify Corni Fructus and its adulterants. METHODS: Genomic DNA extracted from Corni Fructus and its adulterants were used as templates. The ITS (internal trascribed spacer) regions were amplified using polymerase chain reaction. Sequence assembly was performed using CodonCode Aligner V 3.5.4. Genetic distances were computed using MEGA V 5.0. Species identification was conducted using neighbor-joining (NJ) trees. RESULTS: The ITS sequence length of Corni Fructus was 659 bp. The average intra-specific genetic distance of Corni Fructus was 0.005, markedly lower than the inter-specific genetic distance between Corni Fructus and its adulterants (0.357). The ITS2 sequence length of Corni Fructus was 250 bp. No variation was found among the different samples. The interspecific genetic distance of ITS2 between Corni Fructus and its adulterants was 0.571. NJ trees and BLAST results indicated that Corni Fructus and its adulterants can be easily differentiated with monophyly. CONCLUSION ITS/ITS2 regions can accurately and efficiently distinguish Corni Fructus and its adulterants. In addition, the results not only established the foundation for the clinical safety in the utilization of Corni Fructus, but also provided reference for molecular identification of other Chinese herbal medicine and Chinese herbal pieces.
Base Sequence ; Cornus ; classification ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; Molecular Sequence Data ; Molecular Typing ; methods ; Phylogeny ; Species Specificity

Objective To explore the relationship between anxious depression and cortisol rhythm disorder and influencing factors of immune metabolism. And to look for biological markers that can be used for clinical diagnosis and treatment of anxious depression. Methods Totally 43 patients with anxious depres-sion(A-MDD group) and 44 patients with non-anxious depression matched by sex,age and years of education (NA-MDD group)were recruited. Electrochemiluminescence was used to detect the plasma levels of adreno-corticotropic hormone(ACTH),cortisol(COR),c-reactive protein(CRP) and IL-6. Automatic biochemical a-nalysis was used to detect plasma total TC,TG,HDL and LDL. Using logistic regression analysis to discuss the influencing factors of anxiety depression. Results The comparison between the two group showed that the age of first onset,BMI and SBP in the A-MDD group((35. 15±11. 56),(24. 11±3. 03)kg/m2,(130. 09 ±13. 33)mmHg) were significantly higher than those in the NA-MDD group((31. 34± 14. 08),( 22. 70± 3. 19)kg/m2,( 121. 89±12. 49)mmHg)(t=2. 631,2. 009,2. 964,all P<0. 05). The HAMD score and the factor scores of cognitive impairment,change of day and night,delay,sleep disorder and feeling of despair in the A-MDD group((31. 81±5. 39),(8. 03±3. 00),(1. 17±0. 70),(6. 88±1. 93),(4. 44±1. 44),(4. 67± 2. 37)) were significantly higher than those in the NA-MDD group((25. 25±5. 017),(3. 87±3. 12),(0. 79 ±0. 78),(4. 64±2. 22),(3. 34±1. 54),(3. 61±2. 02))(t=2. 297,6. 524,2. 505,5. 210,3. 452,2. 421,all P<0. 05). The plasma TG,CRP and IL-6 levels in the A-MDD group((1. 63±1. 11)mmol/L,(1. 20±0. 77) mg/L,(3. 54±1. 90) pg/L) were significantly higher than those in the NA-MDD group (( 1. 19 ± 0. 66) mmol/L,(0. 933±0. 89)mg/L,(2. 65±1. 34)pg/L) (t=2. 254,2. 250,2. 352,all P<0. 05). The incidence of cortisol disturbance was 72% in the A-MDD group,and 48% in the NA-MDD group,and the difference was statistically significant (χ2=5. 369 P=0. 020). Multivariate Logistic regression found that sleep disorder (β=0. 729,OR=2. 072,95%CI=1. 018-3. 119),IL-6(β=0. 583,OR=1. 792,95%CI=1. 168-2. 748),cog-nitive impairment (β=0. 099,OR=1. 104,95% CI=1. 022-1. 193),cortisol rhythm disorders(β=0. 075, OR=1. 078,95%CI=1. 014-1. 146) were the risk factors for anxious depression. Conclusion Anxious de-pression has a high incidence of cortisol rhythm disorder. The COR and IL-6 may be mediators of cortisol rhythm disorder. IL-6 and cortisol rhythm disorder together with sleep disorder and negative cognition consti-tute maybe high risk factors for anxious depression.