OBJECTIVE: To develop an HPLC method for determination of peoniflorin in Bazhen(eight ingredients) teabag.METHODS:The samples were separated on Diamonsil C18 column(250 mm?4.6 mm,5 ?m) with acetonitrile-water(17∶83) as mobile phase under a detection wavelength of 230 nm.RESULTS: The linear range for peoniflorin was 0.105 3～ 2.106 0 ?g(r=0.999 9) and its average recovery rate was 98.58%(RSD=1.17%,n=9).CONCLUSION: The method is simple,rapid,accurate and applicable for the quality control of Bazhen(eight ingredients) teabag.

Objective To observe the curative effect that losartan potassium combined with enalapril used in treating Ⅳ phase diabetic nephropathy(DN). Methods Divided 74 Ⅳ phase DN suffers into 3 groups randomly.Losartan group:taken losartan potassium lablets 100 mg once every day orally,enalapril group:taken enalapril tablets 10 mg once every day orally,consociation group:taken the two drugs above ai one time with the same dosage.The treatment in the 3 groups were all above 3 years.monitoring the 24-hour albuminuria and SCr back and forth the treatment.Results It worthed statistics meaning that the 24-hour albuminuria in the 3 groups all declined(P＜0.05).The effect of consociation group did better(P＜O.01).In term of SCr,it declined in the consociation group worthed statisties meaning only.Conclusion The consociation application with losartan potassium and enalapril can control the proteinuria of Ⅳ phage DN suffers efficiently as well as defer process of kidney diseases.

Objective To observe the level of autophagy induced by TRAIL in TT cell line and identify the role of autophagy in TRAIL-inducing apoptosis of TT cell line.Methods The growth inhibition of TT cells was measured by MTT assay.MDC staining was used to identify the happening of autophagy.Annexin V/PI double staining was used to analyze the apoptosis rate of TT cells by flowcytometry.The protein expression of caspase-8 and Beclin1 was detected by Western blot.Results ① The growth inhibition ratio of TT cells induced by TRAIL at the concentration of 250,500,1000 and 2000 ng/ml was (3.02 ± 1.82)％,(4.87 ± 1.45)％,(7.51 ± 1.57) ％,(12.76 ± 3.23) ％ respectively,which suggested significant resistance of TT cells to TRAIL.② MDC-labeled green light vesicles was significantly increased after the treatment of TRAIL for 48 h.③ The apoptosis rate of TT cells induced by TRAIL at the concentration of 500 ng/ml and 1000 ng/ml after the pretreat ment of 3-MA for 4 h was(17.83 ± 1.54) ％ and(27.81 ± 1.79) ％ respectively,which was significantly higher than the apoptosis rate induced by TRAIL(3.70 ± 0.34) ％,(6.55 ± 0.59) ％ alone and that induced by 3-MA(7.71 ± 0.64) ％ (t =3.282,P ＜ 0.05 ; t =7.830,P ＜ 0.01).④ The combination treatment of TRAIL and 3-MA increased the cleavage of caspase-8 and down-regulated the expression of Beclin 1.Conclusion Autophagy induced by TRAIL may contributes to the resistance of TT cells to TRAIL,which can be reversed by the inhibition of autophagy.

Objective To investigate the relationship between occult pancreaticobiliary reflux (OPR) and biliary diseases.Methods Forty-four patients with primary biliary diseases was enrolled,and serum amylase level was determined,and the bile in common bile duct (CBD) was collected to measure the amylase level,then the △ amylase was calculated,which equals bile amylase level minus serum amylase level.OPR was confirmed if bile amylase level was higher than serum amylase level,otherwise it would be defined as the control group.Results Among the 44 patients with primary biliary diseases,the incidence of OPR was 72.7％ (n =32).The bile and serum amylase activity of patients with OPR were (1 513 ± 2 725),(44 ± 21)U/L;and they were (18 ± 14) and (38 ± 17) U/L in the control group.The bile amylase level in OPR group was significantly higher than that in the control group,and the difference was statistically significant (P ＜ 0.05),while there was no significant difference in serum amylase activity between the two groups.The incidence of OPR in patients with CBD stones was 100％,and the bile amylase activity was (1 048 ± 1 317) IU/L,and the △ bile amylase activity was (996 ± 1 322) U/L;the incidence of OPR in patients with choledocholithiasis and cholecystolithiasis was 75％,and the bile amylase activity was (2 457 ± 3 312),the △ amylase activity was (2 412 ± 3 320)IU/L;and the corresponding values in patients with gallbladder stones were 80％,(95 ± 82),(57 ± 76)IU/L;and the corresponding values in patients with bile duct cancer were 50％,(73 ± 51),(40 ± 37)U/L.Conclusions The occurrence of OPR is closely related to CBD stones only,CBD stones and gallbladder stones,and it may be one of the main pathogenic factors of bile duct stones.

To investigate the role of Egr-1 in the carcinogcnestic process of hepatocellular carcinoma (H-CC). Methods Expression of Egr-1 gene in HCC tissues were detected by in situ hybridization and immunohistochemistry. Human breast andmouse liver and brain tissues were used for control. ResultsLittle or no Egr-1 transcription was detected both in HCC tissues and in their normal counterparts. High transcription of Egr-1 was detected in the LCD and atrophic-like liver plate of HCC tissues. Protein expression of Egr-1 gene was consistent with mRNA transcription. High expression of Egr-1 protein was also detected in normal breast and mouse brain tissues. ConclusionsLittle or no expression of Egr-1 may play a role in the deregulation of normal growth in the carcinogenestic process of HCC. The differences of Egr-1 expression among liver cells, breast epithelia and mouse brain tissues might be associated with their different ways of proliferation and differentiation in different cell types.

Objective To investigate the inhibition rate of cell proliferation, cell apoptosis rate and their effects on the cell cycle proceeding of the SSMC7721 cell line when 5-FU combined with thermotherapy is induced into the cells, and then provide theoretical bases to the combined therapy of hepatocellular carcinoma. Methods The inhibition rate of cell proliferation was detected by the MTT under different conditions, the cell cycle proceeding and the cell apoptosis rate was detected by flow cytometry and the subcellular structure was detected by the electronmicroscope. Results The cell inhibition rate of the thermotherapy group, 5-FU group and the combinedgroup were 18.4% ,28. 3% and 52. 7% ,respectively. The inhibition rates in the latter two groups were significantly different to the thermotherapy group. The results of flow cytometry showed that the cell numbers increased in G1 stage decreased in S stage,and increased in G2/M stage;the cell apoptosis rate increased. There was significant difference between different groups(P ＜ 0.01 or P ＜0.05). The results of the electronmicroscop showed that the nuclear chromatins agglutinated in the borderline and the mitochondriums became swelled. Conclusions The 5-FU combined with thermotherapy could significantly improve the inhibition rate of cell proliferation, inhibit the cell cycle proceeding from G1 stage to S stage, and induce cells apoptosis and change the subcellular structures in the SSMC7721 cell line.

OBJECTIVEMinimal deviation adenocarcinoma (MDA) is frequently underdiagnosed in pathology tests due to its benign histopathological features. Therefore, its clinical and pathological characters need to be further analyzed and to establish useful immunohistochemical markers to improve the accuracy of pathological diagnosis.

METHODSClinical pathology data were of 13 MDA cases collected and analyzed in the First and Second Hospitals attached to the University, the pathological characters were compared with matched benign hyperplastic and atypical hyperplastic controls of the glandular epithelial cells to find histochemical and immunohistochemical indices valuable for the pathological diagnosis. 8 of these 13 cases were followed and their conditions were discussed.

RESULTSThe main clinical symptoms were watery leucorrhagia, enlargement of the cervix with erosion and hardening. Thickened ligaments were also detected in some cases. The pathological findings included hyperplasia and mild abnormality of the glands, invasion effects into the stroma could be observed in some glands and abortive glands with desmoplastic changes, or edema and inflammatory infiltration around the glands were also observed. The invasion presented in the deep part of the cervix as well, some were found near larger blood vessels or in the small vessels. Compared with benign glands, the immunohistochemical indices of most MDA glands were demonstrated as cancer embryo antigen (CEA) + + +, Ki-67 +--+ + +, P53 +--+ + +, AB/PAS +/+ +, and focal defect on the basement membrane.

CONCLUSIONSOnce any of the diagnostic characters summarized as above was determined, the probability of MDA should be seriously considered, at least atypical hyperplasia in cervical glands must be diagnosed, and follow-up on the patients and histological biopsy of deep tissues (> 5 mm) of the cervix will be necessary to determine the inclusion/exclusion of MDA. Obviously, a correct pathological diagnosis is extremely important for the patient to receive proper treatment in time and thereby improve her quality of life.

Adenocarcinoma ; pathology ; Adult ; Aged ; Cervix Uteri ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Uterine Cervical Neoplasms ; pathology

Objective To study the effect of Pinus yunnanensis on acute alcoholic liver injury in rats and explore its mechanism. Methods A model of acute alcoholic liver injury in mice was prepared by alcohol. The mice were randomly divided into normal control group, model group, positive control group, Pinus yunnanensis low-, medium-and high-dose groups. Mice in the medicine group were given the corresponding medicine by gavage once a day for 7 days. After the last three hours of intragastric administration, the liver and spleen index, ALT, AST and GSH in serum, SOD, MDA and NO in liver homogenates were measured. Histopathological changes of liver were observed by HE staining. Results Compared with model group, Pinecone of Pinus unnanensis high-, medium- and low-dose groups could significantly reduce the liver index in mice (P<0.01), and high dose groups could significantly reduce the number of spleen (P<0.01); The contents of AST in the medium- and high-dose groups significantly decreased (P<0.01) and the GSH activity significantly increased (P<0.05, P<0.01). There was no significant difference in serum ALT level, SOD activity, GSH activity and NO content in the liver tissues of Pinus yunnanensis groups (P>0.05). HE staining results showed that, the damage of liver tissue in mice of Pinus yunnanensis was significantly improved compared with the model group. Conclusion Pinus yunnanensis has protective effects on acute alcoholic liver injury in mice.

Objective To study the effect and related mechanism of celecoxib on tumor necrosis factorrelated apoptosis-inducing ligand(TRAIL) induced apoptosis of medullary thyroid cancer TT cell line.Methods MTT assay was used to measure the growth inhibition induced by TRAIL and celecoxib alone and their combination.TT cell cycle distribution was analyzed by flowcytometry.Hochest33258 staining and DNA ladder was used to detect the apoptosis of drug combination on TT cells.Western blot was used detect the protein change of cyclin A,Cdk2,caspase-8,c-FLIP,and RIP.Results ①MTT showed the growth inhibition ratio of TT cell intervened by the combination of TRAIL and celecoxib was 47.53％ ± 1.34％,which was much higher than that intervened by TRAIL(7.75 ％ ± 3.84％)and celecoxib alone.The differences had statistical significance (t test,F =5.234,P ＜0.01);②PI detection found the cells' number in G0/G1 phase in celecoxib group and combination group were increased compared to that in control group and TRAIL group(F =242.694,P ＜ 0.01);③Western blot indicated the expression of Cyclin A and Cdk2 were down regulated,there was no statistic significance;④ The apoptosis morph in nuclus was detected by Hochest33258 staining and showed the karyopycnosis and muclear fragmentation were increased in combination group with the apoptosis rate 24.23％ ± 2.91％,which was much higher than that in TRAIL(5.86％ ± 1.41％) and celecoxib(20％ ± 1.24％) (t test,F =1.824,P ＜0.01),the difference has statistic significance;⑤Western blot illustrated the active schizolysis of casplase-8 was higher and the expression of c-FLIP and RIP was down regulated in combination group.Conclusion celecoxib plays a positive effect on TRAIL-reduced apoptosis of medullary thyroid cancer TT cell line,which may due to the cell cycle arrest at G0/G1 phase,down-regulation of c-FLIP and RIP and subsequent activation of caspase-8.