To study the expression of HCV non-structure 5 antigen in vitro, a human HepG2 cell line was incubated with a HCV RNA positive serum. The S ABC i mmunological techniques and gold-labeled colloid electron microscopy method wer e employed to examine for the viral proteins in those cells. The HCV non-struct ure 5 antigen was first detected in the HepG2 cells at 72 hours post incubation. The antigen was continuously observed in the cytoplasm or on the membrane as we ll on the cell wall of the HepG2 cells even after 1, 2, 3 and 4 weeks post incub ation. The observation of HCV non-structure 5 antigen continuously expressed in the HepG2 cells strongly indicates that the cells may have been infected by HCV virus and the virus may have replicated in the cells. Therefore, the HepG2 cell line may be served as a potential host for establishment of HCV infection and p ropagation in vitro.

Objective To investigate the features and significance of pathologic changes in apoptosis of small bowel allograft during acute rejection in rats. Methods All 24 recipients were equally divided into four groups ; group A: nonoperative control; group B: allograft ; group C: isograft, group D: treatment control. The graft samples were harvested on day 3, 5, 7, 10 after transplantation, and subjected to histologic examination . Mucosal thickness, villous height and crypt depth were measured, and apoptotic cells of intestinal mucosa of grafts on day 3,5 and 7 after transplantation were examined. Results The mucosal structure was normal in group A; The degree of the inflammatory infiltrated cells ,intestinal mucosa cell apoptosis and structural injury of mucosa in group B were significantly severe compared with groups C and D. As the post-transplanted time increased, the number of musocal apoptotic cells and the degree of mucosal structural injury were significantly increased. The degree of mucosal structural injury in group C was milder than in group B. A few infiltrated cells and mild edema of mucosa occurred in group D , but no mucosal structural injury was found. Conclusions Inflammatory cell infiltration, mucosal epithelial cell apoptosis and mucosal structural damage are the main pathologic features of small bowel allograft during acute rejection. Dynamic observation of the pathologic changes and cell apoptosis of small bowel graft is of certain value in the diagnosis of acute rejection of small bowel graft and in assessment of the degree of small bowel injury.

OBJECTIVETo observe the expressions of Wnt/beta-catenin and the effects of tanshinone IIA (TII A) on Wnt/beta-catenin signaling pathway in high glucose induced renal tubular epithelial cell transdifferentiation.

METHODSHuman kidney proximal tubular epithelial cells (HK-2) were divided into three groups, i. e., the normal glucose group, the high glucose group, and the high glucose plus tanshinone IIA group. The expression of beta-catenin was observed using immunocytochemical staining. The protein expression of beta-catenin, E-cadherin, and alpha-smooth muscle actin (alpha-SMA) were detected by Western blot. The mRNA levels of beta-catenin and E-cadherin were detected by RT-PCR.

RESULTSCompared with the normal glucose group, both the protein and the mRNA expressions of beta-catenin were significantly enhanced (P < 0.01), the expression of E-cadherin significantly decreased (P < 0.01), the expression of beta-catenin increased in the cytoplasm and nucleus in the high glucose group. TIIA at the final concentration of 100 micromol/L significantly reduced the ectopic expression of beta-catenin. At that concentration, the protein and mRNA expressions of beta-catenin in the nucleus significantly decreased, while the protein and mRNA expressions of E-cadherin were up-regulated. Meanwhile, the expression of alpha-SMA obviously decreased.

CONCLUSIONSWnt/beta-catenin signaling pathway participated in the high glucose induced renal tubular epithelial cell transdifferentiation. TIIA inhibited the transdifferentiation process possibly through down-regulating the activities of Wnt/beta-catenin signaling pathway, thus further playing a role in renal protection.

Cadherins ; metabolism ; Cell Line ; Cell Transdifferentiation ; drug effects ; Diterpenes, Abietane ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Glucose ; adverse effects ; Humans ; Kidney Tubules, Proximal ; cytology ; drug effects ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

Objective To study the effects of nitric oxide on Ca2+-activated K+ (KCa) channels in the hypothalamus neurons of newborn SD rats. Methods The data were recorded using inside-out or cell-attached configuration of patch-clamp technique and the kinetic changes of KCa channels are compared before and after 100 μmol/L sodium nitroprusside (SNP) was added into the bathing solution. Results When 100 μmol/L SNP was added to the bathing solution in cell-attached configuration, the open probability of the KCa channels increased from (7.3±1.5)% to (40.2±6.5)%, open time from (7.12±1.41) ms to (15.34±3.45) ms, frequency from (11.3±3.5) Hz to (26.6±4.2) Hz. Conclusion NO-cGMP signal pathway could greatly increase the open probability of the channels as a result of both the prolongation of open period and increase of open frequency, but the pathological or physiological roles it may play require further study.

Objective To study the effects of nitric oxide on Ca2+-activated K+ (KCa) channels in the hypothalamus neurons of newborn SD rats. Methods The data were recorded using inside-out or cell-attached configuration of patch-clamp technique and the kinetic changes of KCa channels are compared before and after 100 μmol/L sodium nitroprusside (SNP) was added into the bathing solution. Results When 100 μmol/L SNP was added to the bathing solution in cell-attached configuration, the open probability of the KCa channels increased from (7.3±1.5)% to (40.2±6.5)%, open time from (7.12±1.41) ms to (15.34±3.45) ms, frequency from (11.3±3.5) Hz to (26.6±4.2) Hz. Conclusion NO-cGMP signal pathway could greatly increase the open probability of the channels as a result of both the prolongation of open period and increase of open frequency, but the pathological or physiological roles it may play require further study.

To evaluate the effects of rhG-CSF and rhSCF on mobilization of the peripheral blood stem cells, 15 monkeys were divided into control, rhG-CSF 10 micro g/(kg x day) and rhG-CSF 10 micro g/(kg x day) + rhSCF 50 micro g/(kg x day) treated groups. Monkeys were administered with vehicle, rhG-CSF and rhG-CSF + rhSCF subcutaneously once daily for 14 days, respectively. The results showed that the highest counts of leukocyte of rhG-CSF treated group were 411% of baseline value on day 7 after administration, compared with that of rhG-CSF + rhSCF treated group which were 538% on day 9. The highest counts of leukocytes lasted for 3 days in combined treated group. CFU-GM from peripheral blood in the two groups were 8.37 and 11.75 times higher at 5 and 9 days respectively after the mobilization. It is concluded that rhG-CSF significantly increases the number of peripheral blood leukocytes and CFU-GM, and a better effect can be obtained by rhSCF + rhG-CSF combined administration.
Animals ; Drug Therapy, Combination ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Hematopoietic Stem Cells ; drug effects ; Leukocyte Count ; Leukocytes ; drug effects ; Macaca mulatta ; Male ; Recombinant Proteins ; pharmacology ; Stem Cell Factor ; administration & dosage ; pharmacology

Postoperative radiotherapy is a major treatment for patients with maxillary sinus carcinoma. However, the irregular resection cavity poses a technical difficulty for this treatment, causing uneven dose distribution to target volumes. In this study, we evaluated the dose distribution to target volumes and normal tissues in postoperative intensity-modulated radiotherapy (IMRT) after placing a water-filled balloon into the resection cavity. Three postoperative patients with advanced maxillary sinus carcinoma were selected in this trial. Water-filled balloons and supporting dental stents were fabricated according to the size of the maxillary resection cavity. Simulation CT scans were performed with or without water-filled balloons, IMRT treatment plans were established, and dose distribution to target volumes and organs at risk were evaluated. Compared to those in the treatment plan without balloons, the dose (D98) delivered to 98% of the gross tumor volume (GTV) increased by 2.1 Gy (P = 0.009), homogeneity index (HI) improved by 2.3% (P = 0.001), and target volume conformity index (TCI) of 68 Gy increased by 18.5% (P = 0.011) in the plan with balloons. Dosimetry endpoints of normal tissues around target regions in both plans were not significantly different (P > 0.05) except for the optic chiasm. In the plan without balloons, 68 Gy high-dose regions did not entirely cover target volumes in the ethmoid sinus, posteromedial wall of the maxillary sinus, or surgical margin of the hard palate. In contrast, 68 Gy high-dose regions entirely covered the GTV in the plan with balloons. These results suggest that placing a water-filled balloon in the resection cavity for postoperative IMRT of maxillary sinus carcinoma can reduce low-dose regions and markedly and simultaneously increase dose homogeneity and conformity of target volumes.
Adult ; Aged ; Carcinoma, Adenoid Cystic ; diagnostic imaging ; radiotherapy ; surgery ; Carcinoma, Squamous Cell ; diagnostic imaging ; radiotherapy ; surgery ; Female ; Humans ; Male ; Maxillary Sinus ; surgery ; Maxillary Sinus Neoplasms ; diagnostic imaging ; radiotherapy ; surgery ; Middle Aged ; Postoperative Period ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Intensity-Modulated ; methods ; Stents ; Tomography, X-Ray Computed

OBJECTIVETo study the specific cellular immune response induced by a recombinant HBV DNA vaccine and evaluate the potential therapeutic effect of this vaccine against HBV.

METHODSA series of HBV epitopes were screened and the optimal combinations of these gene fragments identified to construct the target gene HS which incorporated appropriate flanking sequences. HS was inserted in pVAX1 to construct the recombinant plasmid PVAX-HS. Generation of specific cytotoxic T lymphocytes (CTLs) induced by PVAX-HS in HLA-A2 mice was observed by 51Cr assay and the activity of the CTLs evaluated using ELISPOT assay.

RESULTSPVAX-HS was constructed successfully, which induced specific CTLs with strong activities after immunization in mice as compared with the control group that yielded negative results.

CONCLUSIONPVAX-HS DNA immunization can induced specific cellular immune response in mice, suggesting the potential therapeutic effect of this HBV vaccine.

Animals ; Hepatitis B Vaccines ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Immunization ; methods ; Interferon-gamma ; biosynthesis ; Mice ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; Vaccines, DNA ; genetics ; immunology

OBJECTIVETo investigate the protective effects and mechanism of heat shock response (HSR) on circulatory collapse induced by hyperthermia.

METHODSTwo experiments were carried out: (1) Protective effects of HSR. Rats were divided into 2 groups: heat shock (HS) group, sham control (SC) group. After HS group was pretreated with heat shock and recovered for 20 h at room temperature, both groups were exposed to heat till death, and blood pressure, electrocardiogram were measured continuously during exposure. Mean arterial pressure (MAP), survival time etc were acquired through Chart software. (2) Mechanism of effects. Rats were divided into 3 groups: HS group, SC group and normal control (NC) group. The treatment in HS and SC groups was identical with that in the first experiment, but it would be terminated at 73 min after heat exposure. Systolic pressure (Ps), diastolic pressure (Pd) etc were recorded and content of NO and HSP70 in myocardium were measured.

RESULTS(1) The survival time in HS group [(102.3 +/- 11.4) min] was longer than that in SC group [(87.9 +/- 7.7) min] and shock revealed later (P < 0.01); (2) During early heat exposure MAP in HS group was not different from that in SC group, but after 60 min MAP in HS group were higher than that in SC group; (3) MAP, Ps, Pd, HR and HSP70 in HS group were significantly higher but content of NO was lower than those in SC group (P < 0.01, P < 0.05).

CONCLUSIONHSR may induce upregulation of HSP70 and inhibit excessive production of NO in myocardium, thus result in relief of circulatory collapse induced by hyperthermia.

Animals ; Heat-Shock Proteins ; analysis ; Heat-Shock Response ; physiology ; Hot Temperature ; Male ; Nitric Oxide ; analysis ; Rats ; Rats, Sprague-Dawley ; Shock ; metabolism ; physiopathology ; Time Factors

Objective To develop an autopsy table for the cervical surgery of the small animal to enhance the efficiency by regulating the angles during the operation to expose cervical vertebra adequately.Methods The autopsy table was composed of a table body,a head rack and a foot rack.The head rack was connected with the body by a loose-leaf shaft,which had a screw under a semi-cylindrical cushion put into a sliding slot of the cushion to regulate and fix its position.The table body had four straps at its comers to imobilize the extremities of the experimental animal,and some screws were used to adjust and position the straps to four oblique sliding slots.Results The autopsy table adapted itself to sizes of small animals,which gained advantages in exposure of operating field,short operating time,decreased intraoperative blood loss,head imobilization as well as reduced workload.Conclusion The autopsy table enhances surgery efficiency and safety and has easy operation and low cost,and thus is worthy promoting in the cervical experiment of small animals.