Objective To assess the effect of rhein on diabetic nephropathy in rats with type 2 diabetes mellitus. Methods Diabetes was induced by high-lipids and high-sucrose diets (20% sucrose, 10% lard, 2.5% cholesterol, 1% cholic acid and 66.5 % regular chow, w/w) for 1 month and then intraperitoneal injection with 25 mg/kg streptozoticin(STZ) in female inbred Wistar rats. Rhein was continuously given 100 mg?kg-1?day-1 for 6 months 1 week after STZ injection in rhein-prevented group, and 1 month after STZ injection in rhein-treated group. Blood and urinary biochemistry and renal morphology were evaluated at divers times. Results Rhein ameliorated hypertriglyceridemia and hypercholesterolemia, but had few effect on the level of plasma glucose in both rhein-prevented group and rhein-treated group. Excretion of urinary protein reduced markedly and stable-state plasma glucose (SSPG) level decreased in both groups as compared with diabetic group. Examination of kidney sections from both rhein-prevented group and rhein-treated group showed that the structural lesions of glomeruli including mesangial expansion, diffuse glomerulosclersis and thickening of glomerular basement membrane, as well as artery injury such as arteriolar hyalinosis, atherosclerosis were markedly improved as compared with diabetic group. Morphometry of glomeruli from both rhein-prevented group and rhein-treated group showed that glomerular grow and mesangial hypertrophy occurred in diabetic group were ameliorated. The accumulation of fibronectin examined by immunostaining in glomeruli was reduced in both groups. Conclusion Therapeutic intervention with rhein can halt the progression of diabetic nephropathy and prevent the development of glomerulosclerosis and vascular injuries in this animal model of type 2 diabetes mellitus.

Objective To explore the correlation between membrane targeting of rodent Muc3 C-terminal domain and proteolytic cleavage blockage within its SEA module and N-linked oligosaccharides inhibition.Methods COS-1 cells were transfected with three different expression vectors containing rodent Muc3 C-terminal domain,namely p20,p20t and p20s/a by lipofectAMINE reagent.Inhibition of N-glycosylation of the expressed protein was performed by using tunicamycin.The transfected COS-1 cells(fixed or unfixed) were detected by immunolocalization experiments(anti-V5 and anti-Myc antibody) for the protein expression.Results In fixed COS-1 cells,the expressed product of p20 transfectant detected using both anti-Myc and anti-V5 antibodies was found to localize in perinuclear position and on the plasma membrane.While in the unfixed cells,immunostaining was only confined on cell surface using anti-V5 antibody.The expressed product of p20t transfectant was detected by anti-V5 antibody to localize only in perinuclear region,as observed in a few fixed cells.The distribution of p20s/a fluorescence resembled that of p20 transfectant.Plasma membrane targeting of the non-glycosylated products due to tunicamycin treatment still occurred in transfected COS-1 cells and resembled the glycosylated products.Conclusions The blockage of proteolytic cleavage within C-terminal domain of rodent Muc3 and its inhibition of N-linked oligosaccharides in SEA module cannot affect its membrane targeting.The only apparent requirement for membrane targeting is the transmembrane and/or cytoplasmic tail segments which exist in the C-terminal domains of rMuc3.

Objective To investigate the relation between the expression of HSP70 and p57kip2 in C, CT and Enneking surgical classifi-cation by detecting expression of HSP70 and p57kip2. Methods 30 cases of GCT were collected, including surgical operation specimens and 13 cases of osteochondroma were taken as control group. The expression of HSP70 and p57kip2 were detected, and the relation between the ex-pression of HSP70 and p57kip2 and Enneking surgical classification and prognosis of GCT were analyzed. Results With the increase of tumor Enneking surgical grades, the positive rate of HSP70 was increased and the negative rate of p57kip2 was decreased. Expression rates of HSP70 were 22. 2%, 88.9% ,100% in grades of 1～3 and expression rates of p57kip2 were 88. 9%, 88.9%, 33.3% in grades of 1～3. There was a negative correlation between HSPTo and p.57kip2 expression. Conclusions HSP70 and p57kip2 may play a role in the genesis, and develop-ment of giant cell tumor of bone, and it can be helpful for surgical classification and prognesis of GCT.

Objective: To investigate the relationship between peripheral blood absolute lymphocyte count (ALC) and patients with multiple myeloma (MM). Methods:We obtained clinical features and follow-up data of 102 patients with MM to analyze the prog-nostic value of peripheral blood ALC. Results:Patients with ALC>1.51×109/L were designated as group 1, whereas patients with ALC<1.51×109/L were designated as group 2. Group 1 had significantly lower age as well as concentrations of lactate dehydrogenase (LDH) andβ2-MG than group 2 (P<0.05). The overall survival (OS) of group 1 was significantly higher than group 2 (P<0.05). Sex, ALB, Du-rie-Salmon stage, isotype, subgroup, and stage according to the International Staging System in group 1 were not significantly different compared with group 2 (P>0.05). Conclusion:Lower ALC is a poor prognosis factor. ALC may be an important prognostic factor of MM.

Triptolide (TP) is a major active component in Tripterygium root, but its therapeutic window was very narrow due to its severe multi-organ toxicity. In this work, the effect of TP combined with glycyrrhetic acid (GA) on mRNA expression and activity of four cytochrome P450 (CYP) enzymes in rat liver was studied after intragastric administration of TP (0.05, 0.3 and 0.6 mg x kg(-1) x day(-1)) and TP (0.6 mg x kg(-1) x day(-1)) combined with GA (30 mg x kg(-1) x day(-1)) for 7 consecutive days. Compared with the control, the high dose of TP significantly up-regulated the mRNA expression levels of CYP2E1, 1A2, 3A1 and 2C11, the co-administration of TP and GA further up-regulated the mRNA expression levels of CYP3A1, 2C11 and 2E1 as compared with the high dose of TP. Meanwhile, TP at high dose and combined with GA significantly increased CYP3A-associated testosterone 6beta-hydroxylation activity (2.2-fold and 4.1-fold, respectively) as compared with the control. Because TP is mainly metabolized by CYP3A2 in male rats, the present work indicated that TP-induced increase of CYP3A activity might be an important reason for the rapidly metabolic clearance of TP in rat liver, and GA can reduce the hepatotoxicity of TP by promoting its hepatic metabolic clearance. Furthermore, the results also suggest that the drug interactions might be occurred when TP and GA were co-administered with other CYP3A substrate drug.

Objective:To explore the variation of tumor cell proteins correlated with cell proliferation and chemo-resis-tance after repeated treatment with chemotherapy drugs.Methods:The human lung cancer cell line(LPET-a-2)was repeat-edly treated with chemotherapy drugs:doxorubicin,etoposide,cisplatin and the combination of the3drugs,and each drug was given at2concentrations.The treatment intervals were recorded.ErbB-2,c-myc,MDR1,MRP,LRP,ref-1and NF-?B were tested by flow cytometry.The cells expressing each protein,the mean and total quantity of each protein after each treat-ment were calculated by different drugs at different concentrations.Results:The levels of every protein decreased along with the time of culture.In high-dose group,every item decreased along with the time of treatment.In low-dose group,there was no rule for the item variation with some decreased and some increased.Conclusion:Low-dose anticancer drugs are easier to induce cell proliferation and chemo-resistance than high-dose one,which suggests that adequate chemotherapy should be given in clinical practice.

Objective The C-terminal domain of rodent Muc3 is proteolytically cleaved.This study is to explore the relationship between N-linked oligosaccharides in SEA module and the proteolytic cleavage within C-terminal domain of rodent Muc3.Methods Truncated rodent Muc3 C-terminal domains with complete SEA module(p20SEA) were produced by site-directed mutagenesis to insert a stop code in the required place.Proteins were detected by pulse/chase and immunoprecipitation method,or SDS/PAGE and Western blot.Inhibition of glycosylation of the expressed protein was performed by using tunicamycin.Results Muc3 C-terminal domain was posttranslationally cleaved to produce a V5-tagged 30 000 extracellular glycopeptide and a Myc-tagged 49 000 membrane-associated glycopeptide.Treatment with tunicamycin to transfected COS-1 cells led to the abundant production of 60 000 uncleaved and whole-length Muc3 C-terminal domain,the 30 000 N-terminal fragment shifted to 22 000 and 49 000 C-terminal fragment shifted to 41 000 after deglycosylation.The truncated Muc3 C-terminal domain containing complete SEA module but without the following residues led to production of 36 000 uncleaved and whole-length protein,and 30 000 cleaved product shifted to 22 000 after deglycosylation.Conclusion Proteolytic cleavage in both complete rodent C-terminal domain and complete SEA module without the following residues were partially inhibited by tunicamycin.

Objective To analyze the risk factors of malignant glaucoma for primary angle-closure glaucoma(PACG) patients after surgery.Methods Clinical data of elderly patients with PACG who had received surgical treatment at our hospital between 2013 and 2016 were retrospectively analyzed.Patients were divided into two groups according to whether they suffered (Group A) or not (Group B) from malignant glaucoma after surgery.Results A total of 238 patients (315 eyes) were retrospectively analyzed: 17 cases (22 eyes) in Group A and 221 cases (293 eyes)in Group B.The index of axial length, chamber depth and crystal thickness was lower in Group A than in Group B,and the difference was statistically significant(P<0.05).Univariate analysis showed that age under 50, continuous high intraocular pressure, axial length below 22 mm, chamber depth below 2 mm, crystal thickness below 4.5 mm, completely closed anterior chamber angle and chronic angle-closure type were the possible risk factors for malignant glaucoma after surgery (P<0.05).The logistic regression analysis showed that age under 50 and axial length below 22 mm were independent risk factors for malignant glaucoma for PACG patients after surgery.Conclusion PACG patients,especially those who are young or with short axial length,are more vulnerable to malignant glaucoma.

OBJECTIVE:To observe the effect of electrolytes on the stability of fat emulsion in ALL IN ONE.METHODS: The size of fat emulsion particulates was observed under microscope in 400 - fold magnification.The oil - water demixing was observed with naked eye.RESULTS:The higher the concentration of electrolytes,the stronger the influence on the fat emulsion particulates aggregation and the easier the oil - water demixing appeared.The influence of bivalent ions was stronger than that of monovalent ones.The longer the mixture stored the stronger the influence.CONCLUSION:In order to keep the stability of ALL IN ONE,the concentrations of electrolytes should not be too high,and ALL IN ONE must be used up in 24 hours.

Objective To explore the effect of multiple positioning procedure for prolapse and hemorrhoids (PPH) in treatment of connective tissue type mixed hemorrhoids,and compare the results between PPH and Milligan-Morgan surgery.Methods One hundred and sixteen patients with connective tissue type mixed hemorrhoids were divided into treatment group (62 cases) and control group (54 cases)according to random digits table method.The patients in treatment group received multiple positioning PPH,and the patients in control group received Milligan-Morgan surgery.The operative time,intraoperative blood loss,postoperative pain score,anal margin edema,wound healing time,perianal scar,anorectal stenosis,anal discharge and anorectal resting pressure were evaluated and compared between 2 groups.Results The operative time and intraoperative blood loss in treatment group were significantly lower than those in control group:(15.0 ± 2.3) min vs.(35.0 ± 3.4) min and (5.0 ± 2.1) ml vs.(15.0 ± 3.2) ml,and there were statistical differences (P ＜ 0.05).The postoperative pain score and wound healing time in treatment group were significantly lower than those in control group:(4.83 ± 0.62) scores vs.(7.82 ± 0.37) scores and (8.2 ± 2.6) d vs.(17.4 ± 3.8) d,the rates of anal margin edema,perianal scar and anorectal stenosis in treatment group were significantly lower than those in control group:8.1％ (5/62) vs.37.0％ (20/54),0 vs.29.6％ (16/54) and 1.6％(1/62) vs.16.7％ (9/54),and there were statistical differences (P ＜ 0.05 or ＜ 0.01).The postoperative anorectal resting pressure was significantly lower than preoperative in control group:(13.80 ± 4.20) kPa vs.(17.20 ± 5.87) kPa,and there was statistical difference (P＜ 0.05).One patient (1.6％,1/62) in treatment group occurred anal discharge,and 10 patients (18.5％,10/54) occurred anal discharge in control group.The rate of anal discharge in treatment group was significantly lower than that in control group,and there was statistical difference (P ＜ 0.01).Conclusions Multiple positioning PPH is effective for the treatment of connective tissue type mixed hemorrhoids.It improves the controllability of the amount of removal and the excision site.It decreases the incidence of anal margin edema,perianal scar and anal discharge scar,which provides an effective protection for the anal function.