Aim To observe the protection effect of Tribulus terrestris saponin monomer B (TTSMB) on cadiocytes impaired by hypoxia-reoxygenation (H/R).Methods Cadiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, H/R group,GSTT 100 mg·L~(-1) group and TTSMB 10,1,0.1 nmol·L~(-1) group.Morphocytology change of cadiocytes was observed after the treatment.Cadiocyte survival rate was detected with MTT colorimetric method.Levels of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), malondialdehyde (MDA), and activity of superoxide dismutase (SOD) were determined. Apoptosis rate was detected with flow cytometry.Expression of caspase-3 was examined with western blot.Results Compared with the control group, the survival cell number in H/R group decreased obviously, content of CK,LDH, AST, MAD increased, and activity of SOD decreased (P<0.01).Compared with the model group, the survival cell population increased in TTSMB 10,1,0.1 nmol·L~(-1) group (P<0.05 and P<0.01).Contents of CK, LDH, AST, MAD decreased, whereas the activity of SOD increased (P<0.05 and P<0.01).Apoptosis rate and expression of caspase-3 also reduced in TTSMB 10,1,0.1 nmol·L~(-1) group.Conclusion TTSMB has significant cadiocytes protective effect against H/R and inhibits cadiocyte apoptosis,and the mechanism depends on the effect against oxygen free radical.

Objective To observe the clinical efficacy of Qufeng Qingre Weiling Decoction combined with the decreasing amount of loratadine in treatment of chronic urticaria (CU). Methods Totally 108 CU patients were randomly divided into treatment group and control group, 54 cases in each group. The treatment group was given Qufeng Qingre Weiling Decoction combined with the decreasing amount of loratadine (10 mg, 1 time per day) according to symptom controlling situation, and control group with the decreasing amount of loratadine only. Both groups were treated for 4 treatment courses (2 weeks as a treatment course). Symptom score reduce index was used to calculate the effective rate after 8 weeks of treatment. And the taking antihistamine medicine score was recorded 4, 6, 8 weeks of treatment. The follow-up visits were made to observe long-term efficacy of the cured patients 1 month, 3 months and 6 months after discontinuing the medication. Results Totally 27 cases from the treatment group were cured, with a total short-term effective rate of 96.30%;13 cases from the control group were cured, with a total short-term effective rate of 65.38%, with statistical significance (P<0.05). The taking antihistamine medicine score in treatment group (0.553 ± 0.239) was lower than that in control group (0.721 ± 0.245), namely the symptom improvement in treatment group was better than that in control group by taking less antihistamine medicine. Long-term clinical efficacy showed that the treatment group was also distinctively better than the control group. Conclusion Qufeng Qingre Weiling Decoction combined with the decreasing amount of loratadine is more effective for CU, and with better short-term and long-term clinical efficacy than the control group.

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BACKGROUND: Multipotency of hepatic stem cells is of important value in liver transplantation. Stem cells have been successfully identified and isolated from the animal livers. However, reports on whether stem cells exist in human hepatic tissue and how to isolate and identify them ars few.OBJECTIVE: This study was in attempt to isolate hepatic stem cells from human para-cancerous tissues of hepatoma and in vitro culture them, also to identify the stem cell surface marker, in order to find a new source of heptatic stem cells.DESIGN: Cell observation experiment.SETTING: Department of Common Surgery, First Hospital, Jilin University; Department of Common Surgery,Dongfeng Hospital of Traditional Chinese Medicine.PARTICIPANTS: Samples were harvested from 10 patients with hepatoma admitted to Department of hepatobiliary surgery, First Clinical College, Jilin University between October 2005 and June 2006, with age of 45 to 58 years.Hepatic tissue 2 cm away from cancer nest was cut when patients underwent hepatectomy, and it was pathologically confirmed as carcinoma-free tissue. Written informed consents were obtained from each patient. DMEM/F12 dry powder used for cell culture was provided by Hyclone Company, USA. Fresh fetal bovine serum was prepared by Lianxing Biotech Co.,Ltd, Tianjin. Various cell growth factors were the products of Cytolay Company, USA.METHODS: Para-cancerous tissues of hepatoma was cut into pieces, rinsed with Hank's solution and digested with type Ⅳ collagenase. Then the isolated cells were re-suspended in the DMEM/F12 medium supplemented with 0.1 volume fraction of fetal bovine serum, and hepatocyte growth factors, epidermal growth factors and α- fibroblast growth factors of 25 μg/L each were added in the above medium. When the cultured cells covered 2/3 of bottom,they were digested with trypsinase for passage and inoculated at 2×107 L-1. When cells propagated to the 3rd and 4th generations, 2.60×109 L-1 cell suspension prepared with trypsinase was added, and subsequently, anti-human C-kit antibody, immunomagnetic beads and Buffer solution were added in order. C-kit+ cells were preliminarily isolated by immunomagnetic bead separation. Haematoxylin-eosin staining and immunofluorescent histochemical double-staining were used for detecting the hepatic stem cells in para-cancerous tissues.MAIN OUTCOME MEASURES: ① Observation of cell morphology. ② Identification of hepatic stem cells from para-cancerous tissues. ③ Identification of C-kit+ cells by immunofluorescent histochemical double-staining.RESULTS :① After primarily cultured for 2 weeks, the adherent cells grew in colony. After one half of culture medium was renewed, mature hepatocytes were gradually broken and disappeared. Small round cells propagated, and most of them were located in the center and arranged in cluster. Most cells were found with one big nucleus in each, less cytoplasm and clear cell boundary. When cells propagated to the 1st and 2nd generations, they still grew in colony, but fast. Each C-kit+ cell isolated by immunomagnetic bead separation presented a spherical cell body with a very big nucleus and less cytoplasm. After in vitro cultured for 1 week, it presented broken pieces and apoptotic symptoms.② After para-cancerous tissue was stained by haematoxylin-eosin, atypically proliferated biliary tracts with small round cells could be seen in the portal area. After para-cancerous tissue was stained by immunofluorescent histochemical double-staining, small round cells in the biliary tracts proliferated in the portal arsa co-expressed red fluorescence AFP and green fluorescence cytokeratin (CK) 19 with yellow superposition arsa. ③ After C-kit+ cells were stained by fluorescence immunocytochemisty, cytoplasm expressed alpha-fetoprotein (AFP) red granules and CK19 green granules. The superposition area of both presented yellow fluorescence of AFP+/CK19+-positive cells.CONCLUSION: Hepatic stem cells exist in human para-cancerous tissues of hepatoma. Therefore, expressions of C-kit+/AFP+/CK19+, the surface markers of hepatic stem cells, can be used for identifying and isolating hepatic stem cells. Small round cells obtained by in vitro isolation and culture, i.e. hepatic oval cells possess bipotential differentiation of hepatocyte and hepatobiliary epithelial cells.

Objective To observe the effect of momordicin on tumor necrosis factor-?(TNF-?) level,mRNA transcription,and protein expression in myocardium of viral myocarditis caused by coxsackievirus B3(CVB3),and explore its therapeutic mechanism on viral myocarditis in Balb/c mice.Methods Fifty Balb/c mice were randomly divided into 3 groups as follows:momordicin treatment group(20 cases),vehicle control group(20 cases) and normal control group(n=10).Mice in the vehicle control group and the momordicin treatment group were intraperitoneally inoculated with CVB3,as for the nomal control group,equal amount of culture fluid was given instead.Momordicin[25 mg/(kg?d)] was administered intraperitoneally daily from day 0 to 6.Myocardial histopathology,cardiac TNF-? antigen,protein and mRNA expression were detected on day 15 after CVB3 inoculation,respectively.Results As compared with model group,in mice treated with momordicin,the histological myocardial lesion was significantly reduced [(3.26 ?0.84) vs(1.56?0.48),t=3.90 P

Objective To observe the therapeutic effect of astragaloside on chronic coxsackievirus B3(CVB3)myocarditis in Balb/c mice.Methods Eighty Balb/c mice were randomly divided into 3 groups:astragaloside treatment group(n=30),model of viral myocarditis group(model group)(n=30),and control group(n=20).Mice in the model group and the astragaloside treatment group were monthly intraperitoneally inoculated with CVB3,but equal amount of culture fluid was given instead in control group.The model group and control group were fed with drinking water,astragaloside treatment group were fed with drinking water containing astragaloside at concentration of 300 mg/L for 3 months.Survival rates were determined,myocardial histopathology,collagen volume fraction(CVF) and apoptosis of heart tissue,and CVB3 RNA levels were detected on 3 months later respectively by semiquantitative RT-PCR.Results Compared with model group,in astragaloside treated group,the survival rate on 3 months was significantly improved(59.7% vs 76.7%,?2=4.26 P

Objective To explore the expression of tumor necrosis factor(TNF) ligand-related molecule-1A(TL1A) in mice with viral myocarditis(VM) and the role of astragaloside.Methods Fifty-five male Balb/c mice were randomly divided into 4 groups:control group,model group,low-dose intervention group and high-dose intervention group.Mice in model group,low-dose intervention group and high-dose intervention group were inoculated with 0.1 mL coxsackie B3 virus intraperitoneally.Then,mice in low-dose intervention group and high-dose intervention group were treated with 10 g?L-1 and 90 g?L-1 astragaloside solution,respectively.Mice in control group and model group were treated with 0.1 mL carboxymethycellulose solution.All mice were killed on the 15thday.Histological cross sections of heart were stained with hematoxylin-eosin and myocardial histopathologic scores were counted under optical microscope.The expressions of myocardial TL1A mRNA and protein were detected by reverse transcription polymerase chain reaction and immunohistochemistry.Results The mortality were 0,46.7%,40.0% and 13.3% in control group,model group,low-dose intervention group and high-dose intervention group,respectively.Compared with model group and low-dose intervention group,the mortality was significantly lower in high-dose intervention group(?2=9.46,8.95,Pa

Objective To analyze the pathogenesis,therapy and outcome of pediatric cases with coagulation disorders (CD).Methods All these 137 patients were diagnosed as CD with the methods of hemoglutination five items and/or disseminated intravascular coagulation indexes.Then activity of specific coagulation factors,morphology of bone marrow,hepatorenal function and some other relative tests were performed to find out the cause of CD or the primary disease.Results Forty-three cases were diagnosed as genetic CD with 29 as hemophilia A,4 as hemophilia B and 10 as Von Willebrand disease;while the other 94 patients as acquired CD with 15 as vitamin K-dependence coagulation factor deficiency,22 as hepatic dysfunction,30 as disseminated intravascular coagulation and 1 as thrombotic thrombocytopenic purpura.Genetic CD was treated with replacement therapy to reduce the complication.There was 1 case in this group died of intracranial hemorrhage.Acquired CD was treated with short-term,specific and necessary replacement therapy on the basis of reasonable treatment of primary diseases.Eleven cases died finally in this cohort with 7 cases as liver failure and the other 4 cases as terminal leukemia or lymphoma.Conclusion Pediatric patients with CD were caused by genetic or acquired diseases.In clinic the reason of CD was mainly acquired.The treatment of genetic CD is the replacement of specific coagulation factor for life-long term.The outcome dependes on the lack of degree.While the therapy for acquired CD aims at the primary disease.The principle of blood transfusion is short-term and the outcome dependes on the therapic effects of primary diseases.

This study is to observe the protection of gross saponins of Tribulus terrestris (GSTT) on cardiocytes impaired by adriamycin (ADR) and approach its mechanism of action. Cardiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, model (ADR 2 mg x L(-1)) group, and GSTT (100, 30, and 10 mg x L(-1)) groups. MTT colorimetric method was deployed to detect cardiocyte survival rate, activities of CK, LDH, AST, SOD, MDA and NO were detected, and apoptosis was detected with flow cytometry. Effect of GSTT on caspase-3 was detected with Western blotting. Compared with control group, contents of CK, LDH, AST, MDA and NO were increased, and activity of SOD was reduced (P < 0.05, P < 0.01, P < 0.001) by ADR. Numbers of survival cells were increased (P < 0.05, P < 0.001), contents of CK, LDH, AST, MDA and NO were decreased, and activity of SOD was increased (P < 0.05, P < 0.01, P < 0.001) by GSTT (100 and 30 mg x L(-1)). Apoptosis of cardiocytes and concentration of caspase-3 can be reduced by GSTT (100 and 30 mg x L(-1)). GSTT can protect cardiocytes impaired by ADR, which are possible involved with its effect of resisting oxygen free radical.

Objective To measure the vitamin B1 levels in plasma,erythrocytes and urine of the patients with type 2 diabetes,and to analyze the correlation of vitamin B1 level with the progression of diabetic nephropathy,and to clarify the metabolism of nutrtion mechanism of type 2 diabetic nephropathy.Methods Total 90 patients with type 2 diabetes and 30 healthy volunteers were recruited.According to the levels of microalbuminuria,the patients with type 2 diabetes were divided into non diabetic nephropathy (NDN)group,early diabetic nephropathy (EDN)group and clinical diabetic nephropathy (CDN)group (n=30);the healthy people was used as normal control (NC) group.The vitamin B1 levels in the plasma,erythrocytes and urine were examined with high performance liquid chromatography (HPLC). The correlation of microalbuminuria with the plasma viamin B1 level was analyzed. Results The level of vitamin B1 in plasma of the patients in NC group was (7.1±3.3)μg·L-1,while it was (4.0±2.3)μg·L-1 in NDN group,(3.1±1.0)μg·L-1 in EDN group and (2.3±0.6)μg·L-1 in CDN group. Compared with NC group,the vitamin B1 levels in the plasma in NDN,EDN and CDN groups dropped 43.7%, 56.3%,and 67.6%,respectively (P<0.05). The excretion of vitamin B1 in urine were (2.9 ± 0.8),(9.0 ± 4.7),(11.7±3.9),and (15.6±5.0)μg·L-1 in NC group,NDN group,EDN group and CDN group, respectively.Compared with NC group,the vitamin B1 levels in the urine in NDN,EDN and CDN groups were increased by 2,3 and 4 times,respectively (P<0.05).A negative correlation was found between the level of microalbuminuria and the level of vitamin B1 in plasma (r=-0.62,P=0.013).Conclusion Vitamin B1 deficiency can be observed in the early stage of diabetic nephropathy,and the level of vitamin B1 is closely correlated with the progression of diabetic nephropathy.