Aim To investigate the impact of Cinobu-facini on the proliferation,invasion,and apoptosis of HepG2 cells and the underlying mechanism.Methods The proliferation of HepG2 cells was assessed using the CCK-8 method following treatment with Cinobufaci-ni.The invasion capability of HepG2 cells was evalua-ted through Transwell assay after exposure to Cinobufa-cini.The apoptosis rates of HepG2 cells post Cinobufa-cini intervention were measured using flow cytometry,and the expression levels of VEGF in the culture medi-um of HepG2 cells were determined using enzyme-linked immunoassay.Furthermore,qRT-PCR and Western blot analyses were conducted to assess the im-pact of Cinobufacini on mRNA and protein expression levels related to the CXCL5/FOXD1/VEGF pathway.The interaction between CXCL5 and FOXD1 was inves-tigated via co-immunoprecipitation.Results Cinobufa-cini treatment led to a gradual decrease in HepG2 cell viability in a dose-dependent manner compared to the control group(P＜0.05).Moreover,Cinobufacini sig-nificantly suppressed HepG2 cell invasion(P＜0.05)while enhancing cell apoptosis(P＜0.05).Notably,Cinobufacini exhibited inhibitory effects on the CX-CL5/FOXD1/VEGF pathway,as evidenced by re-duced expression of related mRNA and proteins(P＜0.05).FOXD1 was identified as the binding site of CXCL5.Overexpression of CXCL5 resulted in in-creased proliferation and VEGF secretion by HepG2 cells(P＜0.05),and increased expression of FOXD1 and VEGF(P＜0.05).However,Cinobufacini inter-vention effectively inhibited liver cancer cell prolifera-tion and invasion(P＜0.05),promoted apoptosis(P＜0.05),reduced VEGF secretion by HepG2 cells(P＜0.05),and downregulated the expression of CXCL5 and FOXD1 in HepG2 cells(P＜0.05);but com-pared with the unexpressed group of Cinobufacini,its ability to inhibit cell activity was weakened(P＜0.05),and its ability to inhibit the expression of CX-CL5,FOXD1,and VEGF was weakened(P＜0.05).Conclusion Cinobufacini may inhibit HepG2 cell pro-liferation and invasion and promote HepG2 cell apopto-sis by regulating the CXCL5/FOXD1/VEGF pathway.

OBJECTIVE Bergapten (BG), is a furanocoumarin derived from herbal and citrus extracts can act as antioxidant and selective anticancer agents.The current study aimed to investigate whether bergapten would attenuate immunosenescence and to exploreits immunomodulatory effects on immune responses in D-galactose-induced aging BALB/c mice.METHODS Firstly,mice were given D-galactose(180 mg·kg-1)subcutaneous injections for 30 d.To evaluate the establishment of the aging-related effect in mice, serum samples of BALB/c mice were collected from tail vein. Aging BALB/c mice were freely divided into three groups: negative control group received 1% Tween 80 solution only, named D-gal group. Positive groups were received BG administration at the dose of 20 and 100 mg·kg-1, named D-gal+BG(20)group and D-gal+BG(100)group,respectively.Effects of bergapten on T lympho-cyte proliferation and flow cytometry were assessed by using the splenic cell suspension. Enzyme linked immunospot kits were used to quantitatively determine interferon-γ(IFN-γ)and interleukin-4(IL-4) levels of the isolated serum. Immunophenotype was determined by using mixture of antibodies includ-ing anti-CD3,anti-CD4,and anti-CD8.RESULTS Bergapten(20 mg·kg-1)therapy can modulate immu-nity against viral epidemics and attenuate aging-induced immune deficiency(P<0.01),which was correlat-ed with the decline in the activation of the Th and Tc responses in D-galactose induced aging BALB/c mice.According to the in vivo results,bergapten exposure up-regulated the secretion of IFN-γ and IL-4 in T-helper 1(Th1)and T helper 2(Th2)cells(P<0.05,P<0.01).Additionally,BG(20 mg·kg-1)restored antigen-specific CD4+and CD8+T cells in aging models (P<0.05, P<0.01), which may help to curing chronic infections. CONCLUSION The beneficial effect of bergapten in D-galactose induced aging BALB/c mice may be due to the Th and Tc responses activation.

Objective To explore the effect and mechanism of puerarin on inflammatory response in murine 3T3-L1 adipocytes.Methods Tolllike receptor 4 (TLR4) mediated inflammatory cell models were based on palmitic acid (PA) 200 μmol · L-1 stimulated 3T3-L1 adipocytes cells.And then,these cells were divided into 4 groups:control group(without PA induction in 3T3-L1 cells),model group,low and high dose experimental groups (5,10 μmol · L-1 puerarin).The mRNA expression of TLR4 was detected by Real-time PCR.The tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) were performed using enzyme-linked immuno sorbent assay.The protein expressions of inhibitor of nuclear factor kappa-B kinase β (IKKβ),nuclear factor kappa B inhibitory protein(IκB),nuclear factor kappa B-p65 (NF-κB p65) in 3T3-L1 adipocytes cells were assessed by Western blot.Results The mRNA expression of TLR4 in control group and model group were 1.20 ± 0.20,6.23 ± 0.80;the level of TNF-α in the two groups were (20.40 ± 2.0),(60.20 ± 8.0) pg · mL-1;the level of IL-6 in the two groups were (4.20 ± 0.2),(14.40 ± 0.4) pg · mL-1.Comparison between control group and model group,the diffemce on the three factors had significantly(all P ＜ 0.01).Meanwhile,after administration of puerarin,the protein expressions of IKKβ,IκB and NF-κ-p65 in model group were 2.10 ±0.4,4.20 ±0.5,5.50 ±0.2;that in experimental-H group were 1.40 ± 0.3,2.10 ± 0.4,3.60 ± 0.6.Comparison between experimental-H group and model group,the differnce on the three factors had significantly(all P ＜ 0.01).Conclusion Experimental-H group attenuates palmitate-induced inflammatory response in 3T3-L1 adipocytes,its underlying mechanism include inhibiting expression of TLR4 and downregulating activation of IKKβ,IκB,NF-κB-p65 and decreased expression of TNF-α,IL-6 levels.

Objective To investigate mechanical characteristics of the spirochete flagella with tight-fitting ribbon configuration in micro-periplasmic space. Methods The 2D model of two parallel plates was used to simplify the periplasmic space, and the effects of flagellum spacing and eccentricity on force and torque acted on the spirochete flagella, and wall shear stress acted on the spirochete protoplasmic cylinder were studied by using numerical simulation method. Results (1) The relationship between the flagellum horizontal force and eccentricity was presented as a parabolic curve, and the peak value of the flagellum horizontal force was mainly caused by the gradual increase of pressure difference at two sides of the cylinder and the resistance viscous force as well. Flagellum spacing had no significant influence on flagellum horizontal force. (2) The relationship between the flagellum torque and eccentricity was presented as an exponential curve, and smaller flagella spacing would cause bigger flagella torque. (3) Flagellum spacing had no significant effect on wall shear stress of the protoplasmic cylinder, but it would be increased with the number of flagella and the eccentricity increasing. Conclusions Numerical simulation results in this study can qualitatively reflect mechanical characteristics of the spirochete flagella, and also provide references for further understanding the morphology of spirochete as well as its kinematic mechanism and pathogenic characteristics.

Aim To investigate the effects of predni-sone on trabecular microstructure and biomechanical properties of femur in a rat model of type II collagen-induced arthritis (CIA ) using micro-CT and biome-chanics.Methods Forty 8-week-old male Lewis rats were randomly divided into 2 groups:control (CON ) group with 6 rats,and the remaining 34 rats were used to establish the CIA model.3 weeks after immunization screening CIA rats were randomly divided into CIA group,CIA plus prednisone 4.5 mg · kg-1 · d -1 group and CIA plus prednisone 9 mg · kg-1 · d -1 group.Rats in CON group were given vehicle as well as in CIA group.Rats in the other two groups were treated with prednisone at 4.5 mg·kg-1 ·d -1 or 9 mg ·kg-1 · d -1 .After 90 days treatment,all rats were euthanized,and the left femur was collected for biome-chanics,micro-CT scanning and three-dimensional re-construction.Results Micro-CT data showed that tra-becular thickness,trabecular number,bone volume/total volume,bone mineral density in CIA group were significantly lower than those in CON group.While tra-becular separation,structure model index were signifi-cantly higher than those in CON group.Compared with CON group,biomechanical properties (elastic load, maximum load,break load and stiffness)were signifi-cantly decreased in CIA group.Compared with CIA group,bone volume/total volume and trabecular num-ber were increased,while trabecular separation was significantly decreased in two prednisone groups.Com-pared with CIA group,there was no significant change in biomechanical properties in two prednisone groups. Conclusions Treatment with prednisone for 3 months can ameliorate the damage of trabecular microstructure of the femur in CIA rats,but it has no effect on biome-chanical properties and bone mineral density.

OBJECTIVETo investigate the changes in the expressions of Fas-associated death domain protein (FADD) and cellular-FLICE inhibitory protein (c-FLIP) in the articular cartilage of patients with Kashin-Beck disease (KBD) and the role of these proteins in the pathogenesis of KBD.

METHODSThe cartilage samples were collected from patients with established diagnosis of KBD and osteoarthritis and from healthy control subjects undergoing amputation due to traffic accidents. The expressions of Fas-associated death domain protein (FADD) and cellular-FLICE inhibitory protein (c-FLIP) in the cartilage were detected by immunohistochemistry, and the positive chondrocytes were counted in different layers of the articular cartilage under microscope.

RESULTSThe positivity rates of FADD in the middle layer of articular cartilage from patients with KBD [(28.68∓2.19)%] and osteoarthritis [(35.40∓2.34)%] were significantly higher than that in normal cartilage [(10.51∓5.02)%, F=16.245, P=0.000], but the rates in the upper and deeper layers were comparable among the 3 groups (P=0.206-0.761). In KBD cartilage, FADD expression was the highest in the middle layer [(28.68∓5.38)%] followed by the deeper layer [(17.94∓8.38)%]. Compared with the healthy controls, KBD and osteoarthritis patients showed significantly higher FLIP expression in the upper layer of the cartilage (F=5.929, P=0.018) but similar expressions in middle and deeper layers.

CONCLUSIONSKBD patients have significant increased FADD expression in the middle layer but decreased FLIP expression in the upper layer of the cartilage, suggesting that the death receptor pathway and its regulators play important roles in the pathogenesis of KBD.

CASP8 and FADD-Like Apoptosis Regulating Protein ; metabolism ; Cartilage, Articular ; metabolism ; pathology ; Case-Control Studies ; Fas-Associated Death Domain Protein ; metabolism ; Humans ; Immunohistochemistry ; Kashin-Beck Disease ; metabolism ; pathology

Adult ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; immunology ; Influenza, Human ; prevention & control ; Male ; Middle Aged ; Vaccination

OBJECTIVETo investigate the prognostic implication of common bile duct infiltration in the adenocarcinoma of the ampulla of Vater after panreaticoduodenectomy.

METHODSA retrospective study was conducted on clinical manifestation, pathological behavior and survival data in 102 patients with Vater's ampulla adenocarcinoma, who underwent pancreaticoduodenectomy from Jan 1980 to Dec 2003. The result of patients with the common bile duct infiltration were compared with that of those without.

RESULTSThere were 42 cases in stage I (41.2%), 32 in stage II (31.3%), 27 in stage III (26.5%), and 1 in stage IV (1.0%). As for T stage: 9 cases in stage T1 (8.8%), 40 in T2 (39.2%), 25 in T3 (24.5%), and 28 in T4 (27.5%). As regarding to N stage: 76 cases in stage N0 (74.5%) and 26 in N1 (25.5%). Of these 102 cases, microscopic infiltration in the common bile duct (25.0%) was identified in 26 cases. A significant difference was observed between the patients with bile duct infiltration and those without, in the proportion of pancreatic medullae infiltration: 84.6% (infiltration group) versus 34.2% (non-infiltration group, P < 0.001). Twenty-five cases (24.5%) had recurrence and/or metastases postoperatively, with a median survival of 20 months (range, 2 to 93 months). The overall median survival of the whole group was 46.0 months (2 approximately 192 months), with a significant difference between the common bile duct infiltration group (36 months) and the non-infiltration group (49 months, P = 0.0061). The median non-recurrence survival of the whole group was 43 months (2 approximately 192 months), and a significant difference was observed between the common bile duct infiltration group (35 months) and non-infiltration group (47 months, P = 0.0002).

CONCLUSIONIf the adenocarcinoma of the Vater's ampulla infiltrated the common bile duct, the invasion to the pancreatic medulla is likely developed, and usually with a poor non-recurrence and overall survival. Therefore, postoperative chemotherapy/radiotherapy is suggested.

Adenocarcinoma ; diagnosis ; pathology ; surgery ; Adult ; Aged ; Ampulla of Vater ; Common Bile Duct ; pathology ; Common Bile Duct Neoplasms ; diagnosis ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; secondary ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Pancreaticoduodenectomy ; Retrospective Studies ; Survival Rate

OBJECTIVESTo investigate the clinical-pathological characteristics and surgical prognosis of malignant tumor of pancreatic body and tail.

METHODSA retrospective study was accomplished on clinical manifestation, pathological behavior and postoperative survival in 106 patients with malignant tumor of pancreatic body and tail in single institution from Jan 1980 to Dec 2003, and compared these with 451 patients with malignant pancreatic cancer.

RESULTSThere were significant differences in the following parameters (malignant tumor of the body and tail vs those of the head) between the two tumors: (1) the complaints of pain (0.74:41, chi(2) = 37.035, P < 0.01) and jaundice (0.04:0.75, chi(2) = 155.509, P < 0.01); (2) serum SGPT [(27.33 +/- 3.98) U/L: (118.60 +/- 4.59) U/L, F = 89.351, P < 0.01], total bilirubin [(1.46 +/- 0.46) mg/dl: (14.11 +/- 0.60) mg/dl, F = 105.341, P < 0.01] and albumin [(4.20 +/- 0.45) g/L: (3.91 +/- 0.03) g/L, F = 26.642, P < 0.001]; (3) CEA (0.40:0.24, chi(2) = 6.148, P = 0.046) and CA-19-9 positive rate (0.57:0.86, chi(2) = 24.132, P < 0.01); (4) the concomitant total metastasis (0.38:0.20, chi(2) = 14.266, P < 0.01), including liver metastasis (0.30:0.17, chi(2) = 9.003, P < 0.01). Postoperative median survival, resection of non-metastatic pancreatic body and tail cancer was longer than resection of metastatic disease significantly (15 vs 7 months,chi(2) = 21.63, P < 0.01), which the latter was the same as those who didn't remove (6 months,chi(2) = 0.22, P = 0.64).

CONCLUSIONSThe predominant problem is distant metastasis (especially liver metastasis) in the malignant tumor of the body and tail of the pancreas in comparison with pancreatic head cancer. Resection of the body and tail could not increase postoperative survival if metastasis exists. The major way to improve the prognosis is to prevent and manage the distant metastasis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Pancreas ; pathology ; Pancreatic Neoplasms ; mortality ; pathology ; surgery ; Retrospective Studies ; Survival Analysis ; Survival Rate ; Treatment Outcome

OBJECTIVETo investigate the safety and therapeutic effect of low dose (1000 U/m(2)) L-asparaginase (L-Asp) in the treatment of children with acute lymphoblastic leukemia (ALL).

METHODSSix patients were treated with low dose L-Asp after previously suffered severe side effects from standard dose L-Asp (5000 - 10,000 U/m(2)). Twenty-eight blood samples were obtained randomly from 5 of them. Plasma asparagine concentration was detected by reverse phase-high performance liquid chromatography (RP-HPLC).

RESULTSAll the patients treated with low dose L-Asp showed no any toxic symptoms. The plasma asparagine levels in the patients were all above 5 micromol/L except case 4 (4.91 micromol/L) before receiving L-Asp, and were all decreased below 0.5 micromol/L five days after receiving low dose L-Asp, except case 3 (3.70 micromol/L), the results being like that of receiving standard dose L-Asp.

CONCLUSIONLow dose L-Asp has definite efficacy for childhood ALL, while avoids serious side effects from standard dose L-Asp.

Adolescent ; Antineoplastic Agents ; administration & dosage ; adverse effects ; blood ; Asparaginase ; administration & dosage ; adverse effects ; blood ; Child ; Child, Preschool ; Female ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome