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ObjectiveTo observe the effects of modified Shenqiwan on renal function and fibrosis in diabetic nephropathy mice and explore the underlying mechanism based on the glycogen synthase kinase-3β （GSK-3β）/cyclic adenosine monophosphate （cAMP） response element-binding protein （CREB） signaling pathway. MethodFifty male db/db mice and 10 db/m mice were used in this study. The fifty db/db mice were randomly divided into model group， irbesartan group， and low-， medium-， and high-dose modified Shenqiwan groups. The 10 db/m mice were assigned to the normal group. The mice in the low-， medium-， and high-dose modified Shenqiwan groups were administered with modified Shenqiwan in the dosage form of suspension of Chinese medicinal granules by gavage， those in the irbesartan group were given irbesartan suspension by gavage， and those in the normal and model groups were given distilled water of equal volume by gavage. The intervention lasted for 12 weeks. The blood glucose levels， urine albumin-to-creatinine ratio （UACR）， and the protein expression levels of GSK-3β， CREB， transforming growth factor-β1 （TGF-β1）， E-cadherin， Vimentin， fibronectin （FN）， plasminogen activator inhibitor-1 （PAI-1）， and Collagen type Ⅳ （Coll Ⅳ） in the mouse kidneys were recorded before and after treatment. The extent of renal pathological damage was also observed. ResultCompared with the normal group， the model group showed significant increases in blood glucose levels， UACR levels， and the protein expression levels of GSK-3β， TGF-β1， E-cadherin， Vimentin， FN， PAI-1， and Coll Ⅳ in the kidneys （P<0.05）， decreased protein expression level of CREB （P<0.05）， and severe renal pathological damage. Compared with the model group， the low-， medium-， and high-dose modified Shenqiwan groups and the irbesartan group showed varying degrees of decreases in blood glucose levels， UACR levels， and the protein expression levels of GSK-3β， TGF-β1， E-cadherin， Vimentin， FN， PAI-1， and Coll Ⅳ in the kidneys （P<0.05）， increased expression level of CREB protein （P<0.05）， and improved renal pathological damage. ConclusionModified Shenqiwan can effectively reduce blood glucose levels， improve renal function， and alleviate fibrosis， and the mechanism of action is related to the inhibition of the GSK-3β/CREB signaling pathway.
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Objective:To detect the repair effect of brain protein hydrolyzate(BPH)on the oxidative damage induced by H2O2in the PC12 cells,and to optimize the prescription of BPH stomach floating tablets using the star design effect surface method.Methods:The PC12 cells in the logarithmic phase were divided into normal control group,model group(300 μmol·L-1H2O2),BPH group,artificial gastric juice-treated BPH(GBPH)group, artificial intestinal juice-treated BPH(IBPH)group,artificial gastric juice-treated BPH tablets(GBPH-T)group and artificial gastric juice-treated BPH floating tablets(GBPH-FT)group(The latter five groups were added with 20,40,60,80 and 100 mg·L-1BPH treated with different conditions);at the same time blank control group was set up.The PC12 cells in normal control group didn't receive any treatment,the blank control group was added with medium only,and the PC12 cells in model group were treated with H2O2(300 μmol·L-1)for 3 h. The cell viability was detected by MTT assay.Using Design-expert 8.0.6 Trial software,the dosages of HPMC-K4M,octadecanol and acrylic resin Ⅱ were used as the investigation factors,and the 8 h cumulative release in vitro was used as the evaluation index to optimize the prescription.Results:Compared with normal control group, the activity of PCl2cells in 60 mg·L-1BPH group was increased(P<0.05).Compared with model group,the vitalities of PC12 cells in BPH group,IBPH group,GBPH group and GBPH-FT group were increased significantly (P<0.05 or P<0.01).Compared with BPH group,the cell viability in IBPH group was decreased significantly (P<0.05),and there was no significant difference in GBPH group(P>0.05).Compared with GBPH-T group, the cell viability in GBPH-FT group was significantly increased(P<0.05).The optimal prescription was BPH 20 mg,lactose 10 mg,magnesium stearate 0.4 mg,microcrystalline cellulose 20 mg,HPMC-K4M 27 mg, octadecanol 63 mg,acrylic resinⅡ13 mg;all floating tablets drift in 5 s and sustained floating > 8 h;the difference of the measured value of 8 h cumulative release and the predicted value was not statistically significant (P>0.05).Conclusion:BPH has a significant repair effect on the H2O2-induced oxidative damage in the PC12 cells.Star design-effect surface method has good predictability and reproducibility;it is reasonable and feasible, and can be used to optimize the prescription of BPH stomach floating tablets.
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Objective:To observe the protective effect of deproteinized extract of calf blood (DECB) on the kidney injury of the diabetic rats, and to discuss its mechanism preliminarily.Methods:The male Wistar rats were intraperitoneally injected with STZ (65 mg·kg-1) to establish the diabetes models, then the model rats were randomly divided into model (M)group, and metformin (MMet, 105 mg·kg-1) group , low dose of combined administration (ML,105 mg·k-1 metformin+94.5 mg·kg-1 ,DECB) group, medium dose of combined administration(MM, 105 mg·kg-1 metformin+ 189 mg·kg-1 DECB) group, high dose of combined administration(MH, 105 mg·kg-1 metformin +378 mg·kg-1,DECB) group, another ten Wistar rats were selected as normal control(NC)group.The rats were intragastrically administed with metformin and intraperitoneally injected with DECB, once a day, total of 8 weeks.The rats in NC group and M group were given normal saline solution.The weights and blood glucose levels of the rats in various groups were determined;the levels of serum high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), blood urea nitrogen (BUN), urinary albumin (UAlb), urine creatinine (UCR), serum creatinine (SCr), total cholesterol (TC), triglyceride (TG), uric acid (UA), glutathione (GSH), and malondialdehyde (MDA) were detected;the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined.The pathological changes of kidney tissue of the rats in various groups were detected by HE staining.Results:Compared with NC group, the weight of the rats in M group was reduced(P<0.05),and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG,and MDA were increased(P<0.05);the levels of HDL-C and GSH were obviously reduced(P<0.05),and the activities of SOD adn GSH-Px were obviously reduced(P<0.05).Compared with M group,the weights of the rats in MM and MH groups were increased (P<0.05), and the levels of blood glucose,UAlb,UCr,SCr,UA,BUN,LDL-C,TC,TG, and MDA were decreased (P< 0.05);the levels of HDL-C and GSH were increased (P<0.05),and the activities of SOD and GSH-Px were increased (P<0.05).The pathological observation of kidney tissue showed that the rats in M group had obvios kidney tissue lesions with glomerular congestion and renal tubular edema compared with NC group;compared with M group,the pathological changes of the kidney tissue of the rats in drug administration groups were significantly improved, the renal tubular vacuoles were reduced, and the interstitial hyperplasia was not obvious.Conclusion:DECB combined with metformin can reduce the blood glucose level, regulate blood lipid, improve the pathological changes of kidney tissue in the diabetic rats, reduce the renal damage, and enhance the antioxidant capacity of the body.
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Objective:To observe the protective effect of deproteinized extract of calf blood (DECB)on the ethanol-induced liver injury of the mice,and to preliminaryly discuss its mechanism. Methods:Sixty healthy ICR mice were divided into control group,model group,positive drug group,low,medium and high doses of DECB groups (n=10).By intragastric administration,the mice in control group were given 20 mL·kg-1 saline solution, the mice in low,medium and high doses of DECB groups were administrated with 0.125,0.250,0.500 g·kg -1 DECB,and the mice in positive drug group were administrated with 0.63 g·kg -1 Hugan Tablets;once a day for 30 d. 1 h after the last administration,except control group,the mice in other groups were administrated with one-time grant of 50% ethanol 14 mL·kg -1 ,and fasted for 16 h to establish the models of acute alcohol liver injury.The endurance alcohol time and drunk time of the mice were determined,the activities of aspartate aminotransferase (ALT)and alanine transaminase (AST)activity in serum of the mice were detected,the levels of triglyceride (TG),glutathione (GSH)and malonic dialdehyde (MDA)in liver tissue were determined,and the pathological changes of liver tissue were detected.Results:Compared with model group,the drunk symptoms of the mice in different doses of DECB groups were obviously reduced,the endurance time of the mice in high dose of DECB group and positive drug group was prolonged (P <0.05),and the drinking time was shortened (P <0.05);the ALT and AST activities in serum in mediun and high doses of DECB groups were significantly lower than those in model group (P <0.05).Compared with model group,the MDA and TG levels in liver tissue of the mice in medium and high doses of DECB groups and positive drug group were obviously reduced,and the GSH levels were increased (P <0.05);compared with model group,the pathological damages of liver tissue of the mice in high dose of DECB group caused by ethanol were significantly reduced.Conclusion:DECB can improve ethanol-induced liver injury which may be related to the inhibition of hepatic oxidative stress response.
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@#Objective To investigate the expression of aquaporin (AQP) 1, AQP4, inward rectifying potassium channel 4.1 (Kir4.1) and cytoskeleton features of rat spinal cord astrocytes after cytochalasin D (CytD) intervention. Methods Spinal cord astrocytes isolated from 2~3-day-old rats were cultured till confluency. MTT was used to assess survival rate of astrocytes 2 h, 12 h and 24 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD, respectively. Confocal microscopy was used to observe cytoskeleton features of astrocytes 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml of CytD. The expression of AQP1, AQP4, Kir4.1 mRNA were determined with real-time PCR 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD. Results The survival rate of rat spinal cord astrocytes reduced with the time of co-culture and concentration of CytD (P<0.05). The cytoskeleton of astrocytes was reconstructed. The expression of AQP1, AQP4 and Kir4.1 mRNA increased after co- cultured with 0.05~0.40 μg/ml of CytD. Conclusion The appropriate dosage of CytD may remodel the cytoskeleton and increase the mRNA expression of AQP1, AQP4 and Kir4.1 in spinal cord astrocytes of rats.
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Objective To investigate the expressions of apoptosis stimulating protein of P53 (ASPP)family in colorectal carcinoma tissue and to explore their relationship with the clinicopathological characteristics of colorectal carcinoma,and to clarify the effect of ASPP family in the development of colorectal carcinoma.Methods 45 cases of colorectal carcinoma tissue and 20 cases of healthy controls were selected. Among 45 cases of colorectal carcinoma tissue, there were 1 1 cases of well differentiated colorectal carcinoma, 2 1 cases of moderately differentiated colorectal carcinoma,and 13 cases of poorly differentiated colorectal carcinoma;7 cases of T1 stage colorectal carcinoma,8 cases of T2 stage colorectal carcinoma,25 cases of T3 stage colorectal carcinoma,and 5 cases of T4 stage colorectal carcinoma;19 cases of N1 stage with lymph node metastasis,26 cases of N0 stage without lymph node metatasis.The expressions of ASPP1,ASPP2,and iASPP in 45 cases of colorectal carcinoma tissue and 20 cases of normal colon tissue were detected by immunohitochemistry SP method,and the correlations between the expressions of ASPP family and the pathologic typing,infiltrative depth,and lymph node metastasis of colorectal carcinoma were analyzed. Results ①The immunohitochemical staining results showed that the ASPP family members expressed in colorectal carcinoma tissue and normal colon tissue, and there were no significant differences in ASPP1 and ASPP2 positive rates between colorectal carcinoma tissue and normal colon tissue (P>0.05);the positive expression rate of iASPP in colon cancer tissue was higher than that in normal colon tissue (P0.05);the expression of ASPP2 positive rate was decreased when the differentiation degree of tumor cells reduced,they had positive correlation (rs=0.454,P=0.002);the expression of iASPP in colon cancer tissue had no correlation with the differentiation degree of tumor cells (rs=-0.171,P>0.05).③ The expression of ASPP1 in colon caner tissue had no correlation with the infiltrative depth of tumor (rs=-0.268,P>0.05);the expression of ASPP2 positive rate was decreased when the tumor infiltrative depth increased,they had negative correlation (rs=-0.348,P0.05).④The expressions of ASPP1,ASPP2, and iASPP in colon caner tissue had no correlation with lymph node metastasis (rs=0.089,rs=0.044,rs=0.210, P>0.05).Conclusion The expression levels of iASPP in colon cancer and normal colon tissues are different,it may be useful for the diagnosis, differential diagnosis and evaluation in benign and malignant colorectal diseases. The expression of iASPP is negatively correlated with the pathologic typing and neoplasm staging of colorectal carcinoma,it indicates that iASPP can be used as a indicator in judging the prognosis of colorectal carcinoma.
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Objective To observe effect of muscle basal lamina containing neural stem cells (NSCs) in repair of spinal cord injury.Methods Thirty-six SD rats from the same nest were used in the study and spinal cord hemisection models were induced.The animals were classified to blank control group (clearance of the lesion edge only with isotonic saline),NSCs group (transplantation of NSCs to the edge),NSCs + muscle basal lamina group (transplantation of complex of NSCs and muscle basal lamina to the edge) according to random number table,with 12 rats per group.At weeks 4 and 8,survival and migration of the transplanted cells and compatibility of muscle basal lamina with the host were detected.At weeks 2,4,and 8,the hindlimb function was assayed using BBB scoring system.Results NSCs in NSCs + muscle basal lamina group grew at the lesion edge,migrated to both sides of the edge,and integrated with peripheral tissues.Whereas,few NSCs survived at the lesion edge in NSCs group and inflammatory cell infiltration was notable.At week 2,there was no statistical difference of BBB score among the three groups.At weeks 4 and 8,BBB score in NSCs + muscle basal lamina group (7.92 ± 1.00,11.38 ± 1.51) was significantly higher than that in blank control group (3.82 ± 0.75,3.71 ± 0.76) and NSCs group (6.25 ±1.06,8.25 ± 1.83) (P＜0.05).Conclusion Muscle basal lamina orients growth of NSCs along its lumen,facilitates migration of host cells to ground substance within its lumen,and reduces local inflammatory reaction.
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@#Rheumatoid arthitis is a chronic idiopathic immunodeficient diease and its cause is uncertainty,it is hard to cure radically.Rescently years,the treatment of rheumatoid arthitis with supplementing and boosting the liver and kidney had got great successes.The authors will discuss the following aspects:compound formulas(including classical formula and present formula),traditional Chinese external treatment,treatment of combined traditional Chinese and Western medicine in order to lay a foundation in scientific research in the future.
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Objective:To prepare gACE polyclonal antibody for functional study of gACE.Methods:According to the bioinformatics analysis and prediction of the possible conformational structure,hydrophobicity and antigenicity of gACE and the principal for antibody production,a partial peptide with 18-amino acid residues of gACE was synthesized after homology search.The synthesized peptide was then used to immunize after coupling with KLH.The properties of anti-gACE were analyzed by ELISA,Western blot and immunohistochemistry.Results:The antigenicity was repredicted by bioinformatics analysis.The polyclonal antibody against gACE was successfully obtained and its specificity and sensitivity we conformed by ELISA,Western blot and immunohistochemistry.Conclusion:By the bioinformatics analysis and prediction,the hydrophilicity and antigenicity of gACE are analyzed.The antibody of gACE is successfully obtained.