Adult ; Choristoma ; Female ; Humans ; Nasopharynx ; pathology ; Paranasal Sinus Diseases ; Pituitary Neoplasms ; Sphenoid Sinus ; pathology

Objective To search for the changes of T cells subpopulations and immunoglobulin and their relation-ship in children patients with simple nephrotic syndrome. Design Case-control research. Patients aud Participants 39 patients with simple nephrotic syndrome were divided into two groups:the incipient group and relapse group (6 cases were determined at the incipient and relapse time) .Thereare 28 patients in incipient group, 19 males and 9 females, at the age of 2 to 10 years old. There are20 patients in relapse group, 12 males and 8 females, at the age of 3 to 13 years old. There are 35health children in control group, 21 males and 14 females, 2～13 years old. Interventions T cells subpopulations were determined by indirect immunofluorescence of OKT linesmonoclonal antibodies. The serum IgG was determined by routine simple agar immunodiffusion tests. Results and Conclusions The CD_3~+ and CD_4~+ cells are of no change in the children patients withsimple nephrotic syndrome, and the CD_8~+ and CD_(10)~+ cells are obviously increased, the Values of CD_4~+/CD_8~+ are obviously lower than those in the control qroup, there are no difference between the incipientand relapse groups. The levels of serum IgG were decreased in the 85.3% children patients, IgM were inc-reased in 29.4% of that. The values of CD_4~+/CD_8~+ have positive correlation and negative correlationwith the levels of serum IgG and IgM respectively.

Objective To observe the expression of matrix metalloproteinases(MMP)-1,MMP-3 and iNOS in cartilage of experimental rabbit osteoarthritis at different time intervals after anterior cruciate ligament transection(ACLT)operation.The aim of this study is to provide the theoritical evidence for using ACLT rabbit model in Osteoarthritis(OA)research.Methods Unilateral ACLT was performed on 27 randomly selected while rabbits and underwent unilateral arthrotomy was performed on the other 9 white rabbits as the control group.Nine randomly selected white rabbits in experimental group were killed and 3 white rabbits in the control group at 4th,8th and 12th week respectively.Cartilage degradation of femoral condyles was evaluated macr-oscopically,mRNA expression level and protein expression level of MMP-1,MMP-3 and iNOS was measured by reverse transcription-polymerase chain reaction(RT-PCR)and immunohistochemistry respectively.Results Forepart OA cartilage degradation was observed at the 4th week and became more severe at the 8th week after ACLF.Afterpart cartilage degradation was evident at the 12th week after ACLT while cartilage still remained normal in the control group,mRNA expression level and protein expression level of MMP-1.MMP-3 and iNOS were increased at the 4th week and became higher gradually at the 8th,12th week after ACLT compared with the control group.Expression distribution of MMP-1,MMP-3 and iNOS bad different patterns respectively.Conclusion It is suggested that the process of OA cartilage degradation can be simulated by ACLT model and MMP-1,MMP-3 and iNOS may be good markers in therapeutical research of OA.

Objective To construct the eukaryotic expression vector for the fusion gene comprising the genes encoding mouse anti-human cross-linked fibrin single-chain fragment variable (scFv) antibody and low-molecular weight single-chain urokinase. Methods The single peptide of recombinant human ipro-urokinase was ligated with the target fusion gene, and the same reading frame was guaranteed. The ligated compound was subsequently incorporated respectively into pcDNA3 and pMJK expression vectors by DNA recombination technique. Results and Conclusion The desired recombinant plasmids pcDNA3 and pMJK3 containing the signal peptide of recombinant human ipro-urokinase gene and the fusion gene were constructed,which lays the foundation for establishing of a cell line that may secrete the fusion protein.

Objective To construct the eukaryotic expression vector for the fusion gene comprising the genes encoding mouse anti-human cross-linked fibrin single-chain fragment variable (scFv) antibody and low-molecular weight single-chain urokinase. Methods The single peptide of recombinant human ipro-urokinase was ligated with the target fusion gene, and the same reading frame was guaranteed. The ligated compound was subsequently incorporated respectively into pcDNA3 and pMJK expression vectors by DNA recombination technique. Results and Conclusion The desired recombinant plasmids pcDNA3 and pMJK3 containing the signal peptide of recombinant human ipro-urokinase gene and the fusion gene were constructed,which lays the foundation for establishing of a cell line that may secrete the fusion protein.

Animals ; Bromodeoxyuridine ; metabolism ; Cerebral Cortex ; injuries ; Immunohistochemistry ; Male ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley

AIMTo study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats.

METHODSELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical (alpha-glucosidase activity and carnitine content) changes in different segments of the epididymis were observed.

RESULTSIn the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen. Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosidase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P<0.05).

CONCLUSIONThere were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele.

Animals ; Carnitine ; metabolism ; Epididymis ; enzymology ; pathology ; physiopathology ; ultrastructure ; Male ; Microscopy, Electron ; Rats ; Rats, Sprague-Dawley ; Varicocele ; enzymology ; pathology ; physiopathology ; alpha-Glucosidases ; metabolism

OBJECTIVETo investigate the expression of human epididymal secretary protein 2 isoform human epididymal protein 2beta1(HE2beta1) in the testis and epididymis of adolescent male rats along with its significance.

METHODSImmunohistochemical staining was used to detect the expression and localization of HE2beta1 in the testis and epididymis of 15 adolescent SD rats.

RESULTSHE2beta1 immunoreactive staining was detected in the testis and epididymis. In the epithelia of the epididymal duct, HE2beta1 expressed mainly in the supranuclear region of the principle cells and the basement membrane of some epithelial cells; there were no immunostaining in the n clear cells, halo cells and basal cells. The immunopositive reaction was detected, weak in the distal caput, strong in the proximal, middle corpus and the cauda, but negative in the initial segment. Immunopositive results of HE2beta1 were also observed in some of the nuclei of spermatogonia and Sertoli cells with negatively-stained cytoplasm.

CONCLUSIONImmunohistochemical staining is a fairly sensitive method for detecting HE2beta1 expression. The localization and expression level of HE2beta1 in the genital duct of adolescent male rats exhibited a region- and cell-specific expression pattern, which suggests that HE2beta1 may play an important role in spermatogenesis, maturation and epididymal epithelial innate defense mechanisms.

Animals ; Antigens, Surface ; biosynthesis ; Epididymis ; metabolism ; Glycopeptides ; biosynthesis ; Humans ; Immunohistochemistry ; Leydig Cells ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism

OBJECTIVETo investigate the effects of experimental left varicocele (ELV) on the cystatin-related epididymal spermatogenic (CRES) protein in the testis and epididymis of adolescent rats.

METHODSThe ELV model of Sprague-Dawley (SD) male adolescent rats was established, and the expression of CRES protein in the testis and epididymis was detected by immunohistochemistry and Western-blot at 2 and 4 weeks after surgery.

RESULTSImmunohistochemistry and Western-blot detected CRES protein in both the testis and the epididymis of the ELV rats and the control rats. Immunohistochemistry showed that within the testis, CRES protein was expressed mainly in the cytoplasm of round spermatids and elongating spermatids, sperm acrosomes and residual bodies. The expression was most intensive at Stages I-III and IX-XIV, and then decreased gradually at Stages VII-VII and IV-VI. Within the epididymis, CRES protein was expressed mainly in the cytoplasm of the principal cells of epididymal epithelia. Western-blot detected CRES protein in Mr 19,000 and 14,000, stronger in the former than in the latter. Image and statistical analyses showed that the expression of CRES protein in the 2-week and 4-week ELV groups was significantly higher than in the control group (P < 0.05, or P < 0.01).

CONCLUSIONCRES protein expressed in both the testis and epididymis of adolescent rats and the expression is stage-specific and cell-specific in the testis and segment-specific and cell-specific in the epididymis. The expression of CRES protein in the ELV rats is much stronger than in their corresponding controls. It is suggested that CRES protein may be significantly involved in the regulation of spermatogenesis and sperm maturation, and possibly associated with varicocele-related male infertility or subfertility.

Animals ; Blotting, Western ; Cystatins ; biosynthesis ; Disease Models, Animal ; Epididymis ; metabolism ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism ; Varicocele ; metabolism

OBJECTIVETo construct a recombinant lentivirus vector containing fusion gene NT4-p53(N15)-Ant and transfer it into HepG2 cancer cells for gene therapy.

METHODSThe gene of p53(N15)-Ant was obtained by T-vector method. After restriction enzyme digestion, the interest gene of p53(N15)-Ant was inserted in pBV220/NT4 vector and fusion gene of NT4-p53(N15)-Ant was subcloned into the plasmid of lentivirus and cotransferred into HEK-293 cells with helper plasmid. The recombinant lentivirus was produced by homologous recombination of the above mentioned two plasmids in HEK-293 cells and its titer was measured by plaque-forming. The expression of LV. NT4-p53-Ant in transfected HepG2 cells was finally confirmed by reverse transcription polymerase chain reaction (RT-PCR) procedure. The effect of LV. NT4-p53(N15)-Ant on HepG2 cells was measured by a colorimetric 3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The inhibition effect on HepG2 cells of LV. NT4-p53(N15)-Ant and its potential mechanism was detected by light microscopy, electron microscopy, MTT, LDH-release assay and annexin V-PI double staining.

RESULTSThe gene of p53(N15)-Ant was confirmed by restriction enzyme digestion and DNA sequencing. High titer of the recombinant lentivirus was obtained by homologous recombination in HEK-293 cell lines (1 x 10(11) pfu/ml), and the expression of NT4-p53(N15)-Ant gene in HepG2 cells was confirmed by RT-PCR. The viability of HepG2 cells was decreased to 83.4%, 46.9% and 33.9%, at 24 h, 48 h and 72 h, respectively, after infection by LV. NT4-p53(N15)-Ant. Compared with the LV. EGFP control group, there were significant differences (P < 0.01). The LDH level in HepG2 cells infected by LV. NT4-p53(N15)-Ant at 48 h, 72 h and 96 h after infection was 682 IU/L, 815 IU/L and 979 IU/L, respectively, significantly increased than that in the LV. EGFP group (P < 0.01), indicating the cell membrane destruction.

CONCLUSIONThe recombinant lentivirus vector encoding gene NT4-p53(N15)-Ant is successfully constructed in this experiment by molecular cloning and recombination in vitro techniques, and the results suggested that this fusion gene has an anti-tumor effect, which provides the basis for further research on recombinant adenovirus for cancer gene therapy.

Cell Survival ; Genetic Therapy ; Genetic Vectors ; HEK293 Cells ; Hep G2 Cells ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Lentivirus ; genetics ; metabolism ; Nerve Growth Factors ; genetics ; metabolism ; Nucleotide Transport Proteins ; genetics ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism