Objective:To investigate the effect of upregulated and downregulated PSMA7 on the cell cycle and Cyclin D1,CDK4,P16,Rb of RB pathway in A549 cells.Methods:Transfected upregulated pcDNA3.1-PSMA7 vecter and downregulated pGPU6/Hygro-PSMA7-265 vecter into A549 cells,and then tested the effect of PSMA7 on the cell cycle of A549 cells by flow cytometry,and detected the protein level of Cyclin D1,CDK4,P16,Rb by Western blot.Results:Compared with the control group,the cell cycle of the A549 cells did not change significantly,and the expression of Cyclin D1,CDK4 decreased but P16,Rb increased when PSMA7 was upregulated.Compared with the control group,the proportion of phase G0/G1,G2/M of the A549 cells decreased and phase S increased,and the expression of Cyclin D1,CDK4 increased but P16,Rb decreased when PSMA7 was downregulated.There was statistical significance for those results.Conclusion:PSMA7 could affect the expression of Cyclin D1,CDK4,P16,Rb protein level of RB pathway in A549 and promoted the A549 cells into phase S when it′s downregulated.

Objective To optimize the therapeutic dose and time window of picroside Ⅱ in treating cerebral ischemic injury in rats by orthogonal test. Methods The forebrain ischemia models were established by bilateral common carotid artery occlusion (BCCAO) method. The successful models were randomly grouped according to orthogonal experimental design and treated by injecting picroside Ⅱintraperitoneally at different ischemic time with different doses. The concentrations of MDA, NO and H2O2 in serum and brain tissue were respectively determined by thiobarbituric acid assay, nitratase reductase assay and chemiluminescence immunoassay. Results The optimized composition of the therapeutic dose and time window of picroside Ⅱ in cerebral ischemic injury were ischemia 1.5 h with 10 mg/kg, 1.5 h with 20 mg/kg and 1.5 h with 10 mg/kg body weight according to the expressions of MDA, NO and H2O2 in serum, and ischemia 1.5 h with 10 mg/kg, 1.5 h with 20 mg/kg and 1.5 h with 20 mg/kg body weight according to the expressions of MDA, NO and H2O2 in brain tissue. Conclusion On the basis of the principle of lowest therapeutic dose with longest time window, the optimized composition of the therapeutic dose and time window in cerebral ischemic injury is injecting picroside Ⅱ intraperitoneally with 10-20 mg/kg body weight at ischemia 1.5 h.

In recent years, the prevention and cure of infectious bursal disease (IBD) have become more and more difficult due to the emergence of very virulent strains of infectious bursal disease virus (vvIBDV) and the variant strains of IBDV. In this research, the hybridoma cell lines which secretes anti-idiotypic antibodies against anti-IBDV IgG were established. According to the Jerne's theory of immune network, the use of the anti-idiotypic antibodies as a vaccine will be a new method for the prevention of IBD. In this study, the SPF chickens were inoculated with the IBDV- SD strain, and the bursal was obtained from the died chickens. The bursal was then homogenized and frozen-thawed 3 cycles, and the virus samples were prepared by cane sugar density gradient centrifugation and dialysis. Typical IBDV particles were observed under an electron microscope, and the concentration of the virus protein measured by ultraviolet absorbance spectrophotometry was 10.8 mg/mL. SPF chickens were immunized with the virus and the highly immunized sera were prepared and purified by Sulfuric acid ammonia salt out and Sephadex G-25 chromatography. Then, Balb/C mice of six or eight weeks old were immunized interapertoneally(I. P.) with purified antibodies to IBDV at regular intervals. SP2/0 myeloma cells were fused with the spleencytes from the immunized mice at a ratio of 10:1, in 50% polyethylene glycol (1540) and were then cultured in HAT until all the SP2/0 cells died. The hybridoma cells were selected by ELISA and the highly positive holes were cloned 3 times with the method of limited dilution. Two strains (2B6 strain,5F4 strain) of hybridoma cells were obtained, which were shown by ELISA to steadily secrete anti-IBDV idiotypic antibodies. The chromosome number of the two hybridoma cells were about 88 - 106, 95 in average, and the antibodies secreted belonged to the types of IgG1 and Kappa. Balb/c mice of 3 months old were inoculated I.P. with about 10(7) hybridoma cells per capita, and the ascites were collected 12 days later and the titre of anti-IBDV idiotypic antibodies measured by ELISA was 1 :25600 (for 2B6) and 1:12800 (for 5F4) . The ascites containing the anti-IBDV idiotypic antibodies were emulsified with complete or incomplete Freund's adjuvants, and the anti-IBDV idiotypic antibody vaccine was obtained. SPF and common Jingbai chickens were immunized with the vaccine obtained. The immunized chickens with the vaccine were inoculated with IBDV-SD strain at a dose of 2000 ELD50 after twoimmunizations. All the 10 SPF chickens in the non-immunized group were sick, and 8 of them died; and 5 out of the 50 SPF chickens immunized group got sick and 2 died. All the 10 common Jingbai chickens in the control group were sick, and 6 died; 7 of the 30 immunized common Jingbai chickens got sick and only 1 died. Chi2 analysis showed that the difference between the immunized and the non-immunized groups in both the SPF and the common Jingbai chickens were significant (P < 0.01). Our result indicated that the anti-IBDV idiotypic antibody vaccine well protected chickens and had a great potential in both research and clinical application.
Animals ; Antibodies, Anti-Idiotypic ; biosynthesis ; immunology ; Birnaviridae Infections ; immunology ; Cell Line, Tumor ; Chickens ; Enzyme-Linked Immunosorbent Assay ; Hybridomas ; immunology ; metabolism ; Infectious bursal disease virus ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; Viral Vaccines ; biosynthesis ; immunology

Objective To observe the sequence of fat deposit and its relationship with insulin resistance in SD rats fed by high fat diet.Methods Normal 8-week-old male SD rats were randomly divided into normal chow (NC,n=40)and high fat diet(HF,n=40)groups.Triglyceride(TG)in serum,liver and muscle were measured;glucose infusion rate(GIR)and the mRNA level of genes related to lipid metabolism in liver and muscle were determined in different stages.GIR was detected by eugiyeemic-hyperinsulinemic clamp for evaluating the insulin sensitivity.Gene expression was determined by real-time PCR.Results(1)As compared with NC group,serum TG was not increased after high fat feeding for4 and 8 weeks,it began to increase after 12 weeks [0.52(0.15-1.00) mmol/L vs O.31(0.09-0.53)retool/L, P0.05)in skeletal muscle.After 8 weeks,the expression of ACC1 in liver in HF group was increased by 20.6%,CPT-1 was decreased by 27.1%(P

Objective To evaluate the diagnostic value of MRI in brachial plexus injury.Methods Total 98 patients with brachial plexus injury were examined by MRI before operation.Fifty-four of 98 patients MR imaging were obtained by 0.5 Tesla scanner and other 44 patients were obtained by 1.5 Tesla scanner.The scanning sequences include: SE T_1WI,T_2WI,FFE T_2WI and T_2WI SPIR. Exploration of the supraclavicular plexus was carried out and the MR imaging were compared with the operative finding in 63 patients.Thirty-five patients who had not surgery were followed-up.Results MR imaging found pre-ganglionic injuries in 45 patients and post-ganglionic injuries in 56 patients.Pre-and post-ganglionic injuries simultaneously in 16 patients among them.MR imaging can not find injury sings in 13 patients.The positive rate was 86.73%.MR imaging finding of pre-ganglionic injuries include:(1) Spinal cord edema and hemorrhage,2 patients (4.44% ).(2)Displacement of spinal cord,17 patients (37.78%).(3)Traumatic meningoceles,37 patients (82.22%).(4)Absence of roots in spinal canal, 25 patients(55.56% ).(5)Scarring in the spinal cnanl,24 patients (53.33%).(6)Denervation of erector spine,13 patients (28.89%).MR imaging finding of post-ganglionic injuries include:(1)Trunk thickening with hypointensities in T_2WI,23 patients (41.07%).(2)Nerve trunk complete loss of continuity with disappeared of nerve structure,16 patients (28.57%).(3)Continuity of nerve trunk was well with disappearance of nerve structure,14 patients(25.00%).(4)Traumatic neurofibroma,3 patients (5.36%).Conclusion MR imaging can reveal Pre-and post-ganglionic injuries of brachial plexus simultaneously.MR imaging is able to determine the location (pre-or post-ganglionic)and extent of brachial plexus injury,provided important information for treatment method selection.

Objective To improve recognition and imaging diagnosis of aneurysmal bone cyst secondary to a giant cell tumor.Methods To collect the dates of 12 patients with aneurysmal bone cyst secondary to a giant cell tumor were proved by operation and pathology from January 2003 to October 2006. Analyzed and summarized their imaging manifestations and correlation with pathohistology.Results Six lesions were located in epiphysis and metaphysic regions of long bone.Six lesions were located in pelvis.All cases showed a cystic lesion with expanded and osteolytic,eccentric 10 cases and centric 2 cases.Four cases display trabeculate,the margin is well define with a rim of bone sclerosis in 2 cases.Magnetic resonance imaging(MRI)scans were available in 10 patients.All case showed cystic,dilated lesions with solid areas. Eight cases manifested single or multitude solid nodules in big cystic wall.Two cases appeared solid masses with multitude cysts.The sign of multitude fluid-fluid level,best seen on T_2-weighted images,was present in all patients.Seven cases emerged soft-tissue masses.MR found indicative of large amounts of hemosiderin in one cases.Eight cases were examined by spiral CT with plain scanning and enhancement scanning. Reconstructed image were CTA and 3D-MPR(three dimensions multiplanar reconstruction)imaging.All cases showed cystic,dilated lesions with solid areas.The sign of multitude fluid-fluid level was present in 6 patients.The solid areas and cystic-wall of lesions showed contrast enhancement in 8 patients.3D-MPR imaging showed supply blood vessel of tumors in 3 cases.Arteriovenous malformation did not found in all patients.The surgeons'operative findings and the gross specimens were studied in all patients.All lesions were composed of solid areas and cystic areas.The diagnosis of pathology were ABC with GCT(grade Ⅱ)in 10 cases and ABC with GCT(grade Ⅲ).Conclusion Aneurysmai bone cyst secondary to a giant cell tumor is not rare.Adequately recognizing the pathologic basis of ABC,and selecting imaging techniques correctly (X-ray and MRI,or X-ray and CT)is especially important to diagnose a giant-cell tumor with secondary aneurysmai bone cyst.When an eccentric,expanded,lytic tumor with a cystic-solid lesion in epiphysis of long bone or pelvis shows multiple fluid levels,a giant-cell tumor with secondary aneurysmai bone cyst components should be sufficiently considered.

OBJECTIVETo test the effect of recombinant interleukin-4 (IL-4) and recombinant osteoprotegerin (OPG) in suppressing bone resorption induced by polyethylene wear particles..

METHODSA cranial bone allograft was introduced into the air pouches induced on the back of BALB/c mice, followed by injection of 1 ml suspension of polyethylene particles into the pouches. The mouse models were then divided into 3 groups to receive injections of saline (control), IL-4 alone, or IL-4 and OPG into the pouches. The tissues were harvested 21 days after bone implantation for molecular and histological analyses.

RESULTSPolyethylene wear particles-stimulated inflammatory responses (increased cellular infiltration and IL-1 and TNF production) were markedly reduced by IL-4 treatment either alone or combined with OPG (P<0.05). Polyethylene particles significantly increased tartrate-resistant acid phosphatase (TRAP) staining and bone absorption of the implanted bone graft, and IL-4 treatment, either alone or combined with OPG, obviously reduced the osteolysis induced by polyethylene particles (P<0.05).

CONCLUSIONIL-4 offers protection against polyethylene wear debris-induced inflammation and bone resorption in this mouse model. IL-4 combined with OPG can be a feasible and effective therapeutic approach to the treatment and prevention of polyethylene wear debris-associated osteolysis and aseptic loosening of the prosthetic components.

Animals ; Bone Resorption ; chemically induced ; prevention & control ; Disease Models, Animal ; Female ; Interleukin-4 ; pharmacology ; Mice ; Mice, Inbred BALB C ; Orthopedic Fixation Devices ; adverse effects ; Osteolysis ; chemically induced ; prevention & control ; Osteoprotegerin ; pharmacology ; Polyethylene ; antagonists & inhibitors ; Recombinant Proteins ; pharmacology

Neurogranin, a neuron-specific postsynaptic protein, has been considered to play an important role in synaptic plasticity and learning and memory. The present study aimed to investigate the effects of prenatal restraint stress on neurogranin expression in rat offspring hippocampus. Pregnant rats were given a restraint stress (3 times a day for 7 d, 45 min each time) at the late stage of gestation except that in the control group. The offspring rats were divided into four groups: female control group, male control group, female stress group and male stress group. Expression of neurogranin was determined by immunohistochemistry and Western blot. The results showed that neurogranin-positive immunostaining was detected in all areas of the hippocampus. The staining density was stronger in the CA1 and CA3 regions than that in the dentate gyrus (DG) region. Western blot assay showed that neurogranin protein level in female and male prenatal stressed offspring was significantly lower than that in the controls (P<0.01). Neurogranin level was significantly lower in the female stress group than that in the male stress group, whereas there was no significant gender difference in the control group. Immunohistochemical data further confirmed these results. The present study provides evidence that prenatal restraint stress induces gender-dependent decrease in neurogranin expression in the offspring hippocampus. The prenatal restraint stress-induced decrease in neurogranin expression in the hippocampus might be associated with the deficit in spatial learning and memory reported previously.
Animals ; Blotting, Western ; Female ; Hippocampus ; chemistry ; Immunohistochemistry ; Male ; Neurogranin ; analysis ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley ; Restraint, Physical ; Stress, Psychological ; metabolism

OBJECTIVETo compare the long-term efficacy of procedure for prolapse and hemorrhoids (PPH) and Milligan-Morgan hemorrhoidectomy (MMH) in the treatment of III and IV degree internal hemorrhoids.

METHODSOne hundred patients were randomly divided into two groups and received PPH (n=42) and MMH (n=58) respectively. After two years, the efficacy, complications and function of defecation were compared.

RESULTSTwo years after operation, the morbidities of hydrorrhea (2.38% vs 20.69%, P=0.007), dermal neoplasm formation (9.52% vs 25.86%, P=0.040) and narrowing in the caliber of the stools (2.38% vs 18.97%, P=0.027) were significantly lower in PPH group than those in MMH group (P<0.05). The morbidities of overall complication (9.52% vs 25.86%, P=0.040) and overall abnormal function of defecation (9.52% vs 29.31%, P=0.017) were lower in PPH group than those in MMH group (P<0.05). However, there were no significant differences of the morbidity of relapse (14.29% vs 10.34%, P=0.549), patient satisfactory degree (92.86% vs 87.93%, P=0.636) and overall symptom recurrence rate (19.05% vs 25.86%, P=0.424) between the two groups.

CONCLUSIONSLong-term efficacies of procedure for prolapse and hemorrhoids and Milligan-Morgan haemorrhoidectomy in the treatment of III and IV degree internal hemorrhoids are similar. PPH has better safety, less complications and less effect on abnormal function of defecation compared with MMH.

Adolescent ; Adult ; Aged ; Female ; Hemorrhoids ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Mucous Membrane ; surgery ; Prolapse ; Sutures ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the feasibility and efficacy of a new MRI imaging method--diffusion weighted imaging (DWI) with short TI version recovery-echo planar imaging (STIR-EPI) sequence in differentiating benign cervical lymph nodes from malignant ones. METHODS Twenty nasopharyngeal carcinoma patients and fourteen volunteers received both conventional MRI and DWI with STIR-EPI. Ability of detecting lymph nodes between conventional MRI and STIR-EPI-DWI was compared, and the difference of apparent diffusion coefficient (ADC) value between metastatic lymph node and normal lymph node was analyzed.

RESULTSDWI was more sensitive in detecting lymph node than conventional MRI. ADC value of metastatic lymph node (0. 766 +/- 0. 119) x 10 (-3) mm(2)/s was significantly lower than that of normal lymph node (0. 975 +/- 0. 179) x 10 - mm2/s (P < 0. 01).

CONCLUSIONAs a new MRI imaging technique in detecting cervical lymph nodes, diffusion weighted imaging ( DWI) with short TI version recovery-echo planar imaging ( STIR-EPI) sequence is more reliable and sensitive than conventional MRI imaging, providing an alternative way to differentiate benign lymph nodes from malignant ones.

Adult ; Aged ; Diagnosis, Differential ; Diffusion Magnetic Resonance Imaging ; methods ; Echo-Planar Imaging ; methods ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; Neck ; Reproducibility of Results ; Sensitivity and Specificity