Actins ; metabolism ; Adenomyoepithelioma ; metabolism ; pathology ; surgery ; Aged ; Axilla ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Female ; Humans ; Keratin-7 ; metabolism ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Mastectomy, Modified Radical ; S100 Proteins ; metabolism

Usage of reverse genetic techniques in the research area of the fundamental etiology of foot-and-mouth disease virus (FMDV), has resolved the issue about the function of viral gene of FMDV on genomic integer level. At present, a further recognition and apprehension for the molecular etiology of FMDV based on the development in reverse genetics was made. Combined with the research work in our labs, we reviewed international advances about the molecular pathogenic mechanism, the relationship be-tween virulence and variation in the genomes, influencing factors for the viral replication, and the develop-ment of new-type gene vaccine of FMD in this article, and propose the potential research aspects in reverse genetics of FMDV in the future.

Objective To observe the changes of respiratory and hemodynamic functions in children with different age undergoing laparoscopic Nissen's fundoplication(LNF). Methods Thirty-three children with LNF were divided into three groups according to the age: group Ⅰ,1 to 12 months,n=13;group Ⅱ,1 to 3 years old,n=10;and group Ⅲ,4 to 7 years,n=10.Heart rate(HR),mean arterial pressure,Ppeak,compliance of the respiratory system(CRS) and end-tidal carbon dioxide pressure(PETCO2) were recorded 5 min before pneumoperitoneum(T0),10 min(T1),60 min(T2) after pneumoperitoneum and 10 min after deflation(T3),and parameters of blood gas analysis such as PaCO2 were measured at the same time. Results Compared with those at T0,HR,Ppeak,PETCO2 and the difference between PaCO2 and PETCO2(Pa-ETCO2)were significantly increased at T1 and T2,while CRS was significantly decreased.The most significant changes were found in group Ⅰ. Conclusion The changes of respiratory and hemodynamic functions are observed in children undergoing LNF,and anesthesia management should be enhanced for those within 1 to 12 months old who experience the most significant changes.

Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

Objective To construct phage display single-chain antibody fragments (scFvs) library against histidine-rich protein Ⅱ (HRP-Ⅱ) of Plasmodium falciparum and select specific scFvs of anti- HRP-Ⅱ for the purpose of malaria diagnosis. Method The genes of variable fragments of heavy chain (VH) and light chain (VL) were gained from the spleen cells of BALB/c mice immunized with HRP-Ⅱ protein. The VH and VL genes were then assembled by the method of splicing overlapping extension and cloned into phagemid vector pCANTAB 5E. The scFv phage antibodies were expressed at the surface of the phage after the rescue by helper phage M13K07. HRP- Ⅱ protein was used as antigenic reagent for panning and screening. Results The total RNA from the spleen cells was isolated, and cDNA obtained and VH and VL gene regions amplified using PCR. The VH and VL gene regions were combined with a flexible linker ligated into the pCANTAB 5E phagemid vector, and transformed into TG1 Escherichia coli. The repertoire of the phage antibody was about 106. After panning and screening, 8 positive clones expressed scFv antibodies which were specific for HPR-Ⅱ as demonstrated by ELISA. Conclusion Phage display technology can be used as a powerful tool in making scFv antibodies which have the potential to be used as reagents in the diagnosis and therapy of malaria.

Objective To construct phage display single-chain antibody fragments (scFvs) library against histidine-rich protein Ⅱ (HRP-Ⅱ) of Plasmodium falciparum and select specific scFvs of anti- HRP-Ⅱ for the purpose of malaria diagnosis. Method The genes of variable fragments of heavy chain (VH) and light chain (VL) were gained from the spleen cells of BALB/c mice immunized with HRP-Ⅱ protein. The VH and VL genes were then assembled by the method of splicing overlapping extension and cloned into phagemid vector pCANTAB 5E. The scFv phage antibodies were expressed at the surface of the phage after the rescue by helper phage M13K07. HRP- Ⅱ protein was used as antigenic reagent for panning and screening. Results The total RNA from the spleen cells was isolated, and cDNA obtained and VH and VL gene regions amplified using PCR. The VH and VL gene regions were combined with a flexible linker ligated into the pCANTAB 5E phagemid vector, and transformed into TG1 Escherichia coli. The repertoire of the phage antibody was about 106. After panning and screening, 8 positive clones expressed scFv antibodies which were specific for HPR-Ⅱ as demonstrated by ELISA. Conclusion Phage display technology can be used as a powerful tool in making scFv antibodies which have the potential to be used as reagents in the diagnosis and therapy of malaria.

OBJECTIVETo investigate the influence of Pueraria thomsonii on insulin resistance induced by dexamethasone.

METHOD3T3-L1 adipocytes model of insulin resistance was made by dexamethasone and the change of glucose concentration in cell culture was determined after action of drugs. Rat animal model of insulin resistance was made by intramuscular dexamethasone (1 mg x kg(-1), every other day), and fasting blood glucose (FBG) and fasting serum insulin (FINS) were enamined, and at the end of the experiment, insulin sensitive index (ISI) and insulin resistance index (IRI) were calculated.

RESULTP. thomsonii decreased the concentration of glucose in 3T3-L1 adipocytes culture significantly and improved the sensitivity of 3T3-L1 adipocytes to insulin. P. thomsonii improved the sensitivity of rats to insulin and diminished the fasting serum insulin and insulin resistance index.

CONCLUSIONP. thomsonii can significantly improve insulin resistance induced by dexamethasone.

3T3 Cells ; Adipocytes ; metabolism ; Animals ; Blood Glucose ; metabolism ; Dexamethasone ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Glucose ; metabolism ; Insulin ; blood ; Insulin Resistance ; Male ; Mice ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Rats ; Rats, Wistar

OBJECTIVETo discuss diagnosis and treatment of post traumatic spasmodic flat foot.

METHODSFrom 2002 to 2007, 7 diagnosed patients were recruited and underwent following procedures: under epidural anesthesia or common peroneal nerve block anesthesia, massage was performed on the peroneal muscles for about 5 minutes, then ankle joint was underwent with passive functional exercise. Feeling muscle relaxation, we held the wounded foot in varus and adducted position forcefully, then immobilized it with short leg cast. After 3 weeks of continuous immobilization, the cast was removed and patients were given physical treatment and functional training.

RESULTSSymptoms of these patients such as claudication and pain in foot were improved, the foot deformity was corrected and foot arch restored to normal. After a follow-up of 6 months,no obvious recurrence was observed.

CONCLUSIONThe diagnosis of post traumatic spasmodic flat foot is based on the careful inquiry of traumatic history,physical examination and X-ray results. Manipulative reduction under common peroneal nerve block anesthesia and cast immobilization is a simple and effective method to treat this disease.

Adolescent ; Adult ; Anesthesia ; Female ; Flatfoot ; diagnosis ; etiology ; therapy ; Foot Injuries ; complications ; Humans ; Male ; Middle Aged ; Spasm ; diagnosis ; etiology ; therapy

OBJECTIVEAngiotensin-converting enzyme 2 (ACE-2) has been identified as a functional receptor of severe acute respiratory syndrome coronavirus (SARS-CoV), so its gene was cloned and eukaryotic expressed for further insight into mechanisms in SARS-CoV entry and pathogenesis, as well as development of a safe and reliable neutralization assay for SARS-CoV.

METHODSTotal RNA was extracted from right atrial tissue of a patient with right heart failure resected during a valvular replacement surgery by Trizol one-step method, and the full-length ACE-2 encoding gene was acquired by RT-nested-PCR. The ACE-2 encoding gene was then cloned into pcDNA4/HisMax-TOPO eukaryotic expression vector to construct the recombinant plasmid pcDNA4/ ACE-2, which was then transfected into 293 T cell and ACE-2 eukaryotic transient expression was detected by Western Blot. Syncytia inhibition assay was established to detect SARS-CoV neutralizing antibody, and compared parallelly with SARS pseudovirus neutralization assay.

RESULTSThe recombinant plasmid pcDNA4/ ACE-2 could express ACE-2 protein in eukaryotic cells and induce cell-cell fusion between S protein- and ACE2-expressing cells. This cell-cell fusion assay could be used to detect SARS-CoV neutralizing antibody.

CONCLUSIONSARS-CoV receptor ACE-2 gene was successfully cloned and eukaryotic expressed, and used to establish syncytia inhibition assay for SARS-CoV neutralizing antibody assay.

Animals ; Blotting, Western ; Cell Line ; Cercopithecus aethiops ; Cloning, Molecular ; Gene Expression ; Humans ; Neutralization Tests ; Peptidyl-Dipeptidase A ; genetics ; immunology ; metabolism ; Plasmids ; genetics ; Receptors, Virus ; genetics ; immunology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; immunology ; metabolism ; Transfection ; Vero Cells

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