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1.
Chinese Journal of Epidemiology ; (12): 92-97, 2022.
Article in Chinese | WPRIM | ID: wpr-935355

ABSTRACT

Objective: To evaluate the application of real-time RT-PCR and semi-nested RT-PCR in the detection of norovirus in oysters and analyzing the genetic characteristics of the isolates. Methods: Real-time fluorescent RT-PCR and semi-nested RT-PCR were used to detect norovirus GⅠ/GⅡ in fresh oysters collected from the markets in Beijing from November 2014 to October 2015. The detection rate of the parallel test was also analyzed. In addition, the reliability of semi-nested RT-PCR was evaluated by agreement rate and consistency test (Kappa value). The positive products of norovirus GⅠ/GⅡ capsid protein region gene by semi-nested RT-PCR were sequenced. Software BioEdit 7.0.9.0 was used for sequence alignment, and software Mega 6.0 was used to construct the evolutionary tree. Results: In 72 samples, the detection rate of norovirus was 31.94% (23/72) by real-time RT-PCR, 38.89% (28/72) by semi-nested RT-PCR and 48.61% (35/72) by parallel test. The coincidence rate of the two methods was 73.61%, a moderate degree (Kappa value =0.43). A total of 13 norovirus strains were successfully sequenced, and 11 strains (7 GⅡ.17 strains, 2 GⅡ. 4 Sydney_ 2012 strains, 1 GⅡ. 1 strain and 1 GⅡ. 21 strain) were obtained from norovirus positive samples by two RT-PCR methods, two strains (1 GⅡ. 17 strain and 1 GⅡ. 3 strain) were obtained from real-time RT-PCR negative samples which were positive for norovirus by semi-nested RT-PCR. The similarity between these strains and reference strains from diarrhea patients, environmental sewage, and shellfish products were 84.4% - 100.0%. Conclusions: The parallel test of norovirus in oysters by two RT-PCR methods can improve the detection rate and detect more genotypes. Norovirus strains in oysters were highly homologous with reference strains from diarrheal patients, environmental sewage, and shellfish products. Therefore, surveillance, prevention and control for norovirus should be carried out in people who have frequent contacts with oysters and related environments.


Subject(s)
Animals , Humans , Beijing , Norovirus/genetics , Ostreidae , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction
2.
National Journal of Andrology ; (12): 291-295, 2012.
Article in Chinese | WPRIM | ID: wpr-238964

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the outcomes of perineal urethrostomy plus secondary urethroplasty for ultralong urethral stricture and assess its influence on the patient's quality of life.</p><p><b>METHODS</b>We retrospectively analyzed 54 cases of ultralong urethral stricture treated by perineal urethrostomy from 2000 to 2010. The mean age of the patients was 40 years, and the average length of stricture was 6.5 cm. We evaluated the patients'quality of life by questionnaire investigation and the clinical outcomes based on IPSS, Qmax, the necessity of urethral dilation and satisfaction of the patients.</p><p><b>RESULTS</b>The mean Qmax of the 54 patients was (14.0 +/- 4.7) ml/min. Of the 34 cases that underwent secondary urethroplasty, 22 (64.7%) achieved a mean Qmax of (12.0 +/- 3.5) ml/min, 8 (23.5%) needed regular urethral dilatation and 4 (11.8%) received internal urethrotomy because of restenosis. IPSS scores were 5.4 +/- 2.1 and 8.5 +/- 5.8 after perineal urethrostomy and secondary urethroplasty, respectively. Fifty of the total number of patients (92.6%) were satisfied with the results of perineal urethrostomy, and 22 of the 34 (64.7%) with the results of secondary urethroplasty.</p><p><b>CONCLUSION</b>Perineal urethrostomy plus secondary urethroplasty is safe and effective for ultralong urethral stricture, and affects very little the patient's quality of life.</p>


Subject(s)
Adolescent , Adult , Aged , Child , Humans , Male , Middle Aged , Young Adult , Ostomy , Perineum , General Surgery , Quality of Life , Retrospective Studies , Treatment Outcome , Urethral Stricture , General Surgery , Urologic Surgical Procedures , Methods
3.
Chinese Journal of Surgery ; (12): 1781-1784, 2010.
Article in Chinese | WPRIM | ID: wpr-346383

ABSTRACT

<p><b>OBJECTIVE</b>To study the neurophysiologic of detrusor overactivity (DO) due to partial bladder outflow obstruction (PBOO).</p><p><b>METHODS</b>Twenty four female Wistar rats with DO caused by PBOO were studied simultaneously with ten sham-operated rats. An electrophysiological multi-channel simultaneous recording system was used to record pelvic afferent fiber potentials as well as the pudendal nerve motor branch potentials, external urethral sphincter electromyogram (EUS EMG) and abdominal muscle EMG during filling cystometry. To test the effect of the unstable contraction in DO rats after the decentralization of the central nervous system, DO rats were studied the changes of the unstable contraction after transection of the spinal cord (T(8) level), pelvic nerve, the sympathetic trunk, and the pudendal nerve.</p><p><b>RESULTS</b>The incidence of DO was 62.5% in filling cystometry. During filling cystometry, there are two type of DO contraction according to the changes of pelvic afferent fiber signals, the relevant nerves and muscles responses: the small pressure of the unstable contraction (S-DO) and the big pressure of the unstable contraction (B-DO). For the B-DO, there were significant changes in the recordings of pelvic afferent fiber, the motor branch of the pudendal nerve, EUS EMG, and abdominal muscle EMG. While all these differences have not been recorded during S-DO. Both the filling-voiding cycle and the unstable contraction of B-DO were eliminated and the base line of bladder pressure increased after T(8) spinal cord transection. While the S-DO was not affected by such transection. When bladder relevant nerves were transected by the sequence of the pelvic nerve, the sympathetic trunk, and the pudendal nerve, the filling-voiding cycle was eliminated. The base line of bladder pressure increased significantly. No B-DO was recorded, but the S-DO still existed.</p><p><b>CONCLUSION</b>There are some bladder-genic factors take part in the DO contractions induced by PBOO.</p>


Subject(s)
Animals , Female , Rats , Disease Models, Animal , Pelvic Floor , Rats, Wistar , Urinary Bladder , Urinary Bladder Neck Obstruction , Urinary Bladder, Overactive
4.
Chinese Journal of Gastrointestinal Surgery ; (12): 364-367, 2009.
Article in Chinese | WPRIM | ID: wpr-326497

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate anorectal dynamics, function and efficacy of ultralow rectal carcinoma patients undergone intersphincteric resection(ISR).</p><p><b>METHODS</b>From January 2004 to August 2007, 30 patients with ultralow rectal carcinoma(2.5-4.0 cm distance from anal edge) underwent ISR. All the patients received anorectal manometry before and after operation. The postoperative anal function was evaluated by Williams continence standard and the treatment outcome was followed up.</p><p><b>RESULTS</b>After ISR operation, anal resting pressure, maximum squeeze pressure and maximum tolerance volume of the rectum decreased significantly (all P<0.01) and restored gradually, but not to normal. The rectal anal inhibitory reflex disappeared in 27 patients(90.0%) and was not improved. According to Williams continence standard, 86.7%, 93.3% and 96.7% of patients obtained acceptable anal function in 3, 6, and 12 months after operation respectively. During follow-up of 12 to 44 months, all the patients were still alive and no patient developed pelvis or local recurrence, distant metastasis and anastomotic leakage. Fecal eczema of anus occurred in 10 patients, colonic mucosa prolapse in 2 patients and stenosis of anal canal in one patient.</p><p><b>CONCLUSION</b>ISR for ultralow rectal carcinoma can not only attain radical treatment outcome, but also preserve anal sphincter.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Anal Canal , General Surgery , Anastomosis, Surgical , Methods , Digestive System Surgical Procedures , Rectal Neoplasms , General Surgery , Rectum , General Surgery , Treatment Outcome
5.
National Journal of Andrology ; (12): 1007-1013, 2009.
Article in Chinese | WPRIM | ID: wpr-252841

ABSTRACT

<p><b>OBJECTIVE</b>To determine the effects of cigarette smoking on the cyclogeny of spermatogenic cells in rats.</p><p><b>METHODS</b>Rat models of passive smoking were established using a self-made smoking device, and then allocated randomly into two passive smoking groups (A and B, n = 10) and two corresponding control groups (C and D, n = 10). Groups A and B were exposed to cigarette smoke for 8 weeks, followed by the sacrifice of the rats in Groups A and C. And the animals in Groups B and D were killed 48 days after the cessation of passive smoking. The spermatogenesis cycle of each group of rats was detected by flow cytometry, the levels of testosterone (T) and luteinizing hormone (LH) measured by radio-immunity method, and the testis histopathology analyzed by HE staining and transmission electron microscopy.</p><p><b>RESULTS</b>Compared with Group C, Group A showed a significant decrease in the number of spermatids, spermatozoa ([18.76 +/- 3.58]%) and primary spermatocytes ([5.71 +/- 1.18]%) (P < 0.01), but an obvious increase in the spermatogonias ([55.98 +/- 5.35]%, P < 0.01), with a markedly decreased proliferation index ( P < 0.01). The rats of Group A also exhibited pycnosis of spermatocytes, nucleus aberration of Leydig cells, expansion and degranulation of the endoplasmic reticulum, decreased Golgi apparatus, increased lysosomes and fat drops of Sertoli cells, as well as a reduction in the thickness of the wall and the layers of seminiferous tubules and the number of spermatogonia. The T and LH levels were significantly lower in Group A than in C (P < 0.01). After the cessation of passive smoking, a remarkable increase was observed in the percentage of spermatozoa and primary spermatocytes and the levels of serum T and LH in Group B, although the latter were still lower than those of Group D.</p><p><b>CONCLUSION</b>Smoking damages spermatogenic epithelia, Leydig cells and Sertoli cells, reduces the T and LH levels, and block the proliferation of spermatogenetic cells. These changes can be partially reversed after cessation of smoking.</p>


Subject(s)
Animals , Male , Rats , Rats, Wistar , Smoking , Spermatogenesis , Testis , Pathology
6.
Chinese Journal of Surgery ; (12): 1542-1545, 2008.
Article in Chinese | WPRIM | ID: wpr-258328

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of ICC-like cells in bladder neuromodulation in rat urinary bladder.</p><p><b>METHODS</b>14 SD rats and 1 guinea pig were sacrificed in this study. The ultra structural relationships among interstitial cells, nerves and detrusor smooth muscle cells (DSMCs) of urinary bladder were investigated by transmission electron microscopy (TEM). c-kit immunofluorescence was used to identify ICC-like cells in SD rat urinary bladder and the structural relationship between ICC-like cells and nerve terminals was studied by immunofluorescence (double-label).</p><p><b>RESULTS</b>Gap junction between ICC-like cells and DSMCs was confirmed by TEM. ICC-like cells were very close apposition with nerve terminals under TEM. ICC-like cells were identified to exist in sub-urothelium layer, along the longitude of smooth muscle bundles and among detrusor smooth muscle in SD rat urinary bladder by c-kit immunofluorescence. Double-labeled tissue with c-kit and PGP9.5 antibodies also showed that ICC-like cells were very close apposition with nerve terminals in SD rat bladder.</p><p><b>CONCLUSIONS</b>Morphological study indicated that ICC-like cells in rat urinary bladder may play an important role in detrusor neuromodulation. Further study on function will be helpful for elucidating the mechanism of bladder neuromodulation clearly.</p>


Subject(s)
Animals , Female , Male , Rats , Cells, Cultured , Gap Junctions , Guinea Pigs , Muscle, Smooth , Myocytes, Smooth Muscle , Cell Biology , Nerve Endings , Rats, Sprague-Dawley , Urinary Bladder , Cell Biology
7.
Chinese Medical Sciences Journal ; (4): 192-195, 2007.
Article in English | WPRIM | ID: wpr-243530

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of alcohol intake on penile structure and function in rats.</p><p><b>METHODS</b>Thirty adult male Wistar rats were randomly divided into two groups: control group and alcohol intake group. They were administered with 2 mL of normal saline and 40% alcohol solution respectively through gastric tubes every day. Three months later, the animal model of alcohol intake was evaluated by modified Nayagida's method, and the effects of alcohol on the rats were studied by sexual behavior, the number of apomorphine-induced penile erection, level of testosterone in the sera, and the content of penile smooth muscle.</p><p><b>RESULTS</b>The scores of animal model of alcohol intake evaluated by Nayagida's method were 0.66 +/- 2.05 in the control group and 9.26 +/- 5.50 in the alcohol intake group (P < 0.05), which indicated that an animal model of alcohol intake was successfully established. Sexual behavior, the number of apomorphine-induced penile erection, testosterone level in the sera, and the content of penile smooth muscle of the alcohol intake group were all statistically different as compared with the control group (P < 0.05).</p><p><b>CONCLUSION</b>Alcohol intake induces sexual dysfunction in rats, which may be due to the decline of testosterone level in the sera and decline of penile smooth muscle.</p>


Subject(s)
Animals , Female , Male , Rats , Ethanol , Penis , Physiology , Rats, Wistar , Sexual Behavior, Animal , Testosterone , Blood
8.
Chinese Journal of Burns ; (6): 409-412, 2007.
Article in Chinese | WPRIM | ID: wpr-347665

ABSTRACT

<p><b>OBJECTIVE</b>To investigate whether the third-party dendritic cells (DC) incubated with donor's antigens possess the similar immune functions with donor derived immature DC.</p><p><b>METHODS</b>Female C57 BL/6, BALB/c and Kunming mice were used as skin transplant donors, recipients and third-party, respectively. Forty BALB/c mice were randomly divided into A (normal control), B (cyclophosphamide administration), C (CTX and donor derived immature DC), D (CTX and third-party immature DC), E (CTX and third-party DC loaded with donor's antigens) groups, with 8 mice in each group. The mice in group B, C, D and E were given CTX (200 mg/Kg) 4 days before operation, while those in group A were given equivalent amount of normal saline(NS). Then in group C, D and E, DC at a dose of 1 x 10(7)/ml were intraperitoneally injected with 2 days before grafting and 12 days after operation, but the mice in group A and B were given NS in the same manner. Mean survival time (MST) of skin grafts was recorded, and biopsies of grafts on 5 and 10 post-operation days (POD) were harvested for histologic examination.</p><p><b>RESULTS</b>Compared with group A [(16.1 +/- 3.5)d], MST of skin grafts in group C [(38.3 +/- 7.7) d] and E [(34.9 +/- 7.7) d] were significantly prolonged ( P < 0.01), while no obvious difference was observed between group C and E( P > 0.05), but there was statistically significant difference in MST between group D [(23.7 +/- 2.7) d] and E ( P < 0.05). In addition, clear epithelial structure and infiltration of inflammatory cells were observed in specimens from both groups C and E.</p><p><b>CONCLUSION</b>Both donor derived immature DC and third-party DC loaded with donor's antigens can partly induce donor specific transplantation tolerance.</p>


Subject(s)
Animals , Female , Mice , Cyclophosphamide , Pharmacology , Dendritic Cells , Allergy and Immunology , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred Strains , Random Allocation , Skin Transplantation , Allergy and Immunology , Transplantation Tolerance , Allergy and Immunology , Transplantation, Heterologous , Allergy and Immunology
9.
National Journal of Andrology ; (12): 1099-1101, 2006.
Article in Chinese | WPRIM | ID: wpr-289070

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the relationship between the intake of ethanol and sexual function of rats.</p><p><b>METHODS</b>Sixty adult male Wistar rats were randomly divided into five groups: the control, 10% , 20% , 30% and 40 ethanol groups, which received. . 9% sodium chloride, 10% , 20% , 30% and 40% ethanol solutions respectively at a dose of 2 ml through gastric tubes once a day. Three months later, we observed the effects of ethanol on the sexual function of the rats by their sexual behaviors, the number of apomorphine-induced penile erections, and the content of testosterone in the serum and nitric oxide synthase ( NOS) in the penis.</p><p><b>RESULTS</b>Compared with the control group, the number of apomorphine-induced penile erections in the 10% and 20% ethanol groups was not inhibited significantly (P > 0.05), but the latent period of mounting and intromission in the 10% ethanol group was prolonged and the sexual behaviors in the 20% ethanol group were inhibited except the latent period of ejaculation. The sexual behaviors and the number of apomorphine-induced penile erections of the 30% and 40% ethanol groups were inhibited significantly (P < 0.05). Testosterone in the serum and NOS activity in the penis of the experimental groups were reduced (Pat < 0.05).</p><p><b>CONCLUSION</b>An adequate volume of ethanol does not induce sexual dysfunction in rats, but long term and excessive intake of ethanol may cause penile erectile dysfunction.</p>


Subject(s)
Animals , Male , Rats , Dose-Response Relationship, Drug , Ethanol , Pharmacology , Nitric Oxide Synthase , Metabolism , Penile Erection , Penis , Metabolism , Random Allocation , Rats, Wistar , Sexual Behavior, Animal , Testosterone , Blood
10.
Chinese Journal of Burns ; (6): 203-206, 2006.
Article in Chinese | WPRIM | ID: wpr-312493

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the influence of liposome induced gene transfection on the phenotypic characteristics and immune function of human immature dedritic cells (imDC).</p><p><b>METHODS</b>Monocytes were isolated from human cord blood, and they were differentiated into imDC by rhGM-CSF and rhIL-4 induction. Then the morphologic observation and immune phenotypic identification were performed in im-DCs. imDCs were divided into transfection (T, with liposome transfection of pEGFP vector) and control (C, without transfection) groups. The transfection rate and expression of cell maturation marker (CD83, CD86 and HLA-DR) were determined with flow cytometry, and the proliferation of non-sensitized T lymphocyte before and after transfection was determined with allogeneic mixed leukocyte reaction (MLR).</p><p><b>RESULTS</b>im-DC derived from human cord blood cells had typical appearance and surface markers consistent to what reported in the literature. The expression rates of CD86, CD83 and LA-DR in T group were (12 +/- 6) %, (8.6 +/- 2.3) % and (71 +/- 7) %, respectively, which exhibited no difference compared with those in C group (13 +/- 6) %, (9.1 +/- 3.8) % and (72 +/- 8) %, (P > 0.05). MLR results indicated that there was no obvious change in the immune stimulation function of imDC after transfection (P > 0.05) , with stimulation index lower than 2.</p><p><b>CONCLUSION</b>There is no change in maturation of imDC after liposome transfection, but the transfection efficiency needs to be elevated.</p>


Subject(s)
Humans , Antigens, CD , Metabolism , B7-2 Antigen , Metabolism , Cell Separation , Dendritic Cells , Cell Biology , Allergy and Immunology , HLA-DR Antigens , Metabolism , Immunoglobulins , Metabolism , In Vitro Techniques , Liposomes , Metabolism , Lymphocyte Activation , Membrane Glycoproteins , Metabolism , T-Lymphocytes , Allergy and Immunology , Transfection
11.
Chinese Journal of Burns ; (6): 211-214, 2006.
Article in Chinese | WPRIM | ID: wpr-312491

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the influence of low doses of granulocyte macrophage colony stimulating factor on the allogeneic antigen (Ag) ingestion capacity of immature dendritic cells (GM low DC), and subsequently the changes in the cellular phenotype and function.</p><p><b>METHODS</b>Mononuclear cells from C57BL/6 mice was labelled with 3H-Leu to make Ag supernatant. The Ag supernatant was cocultured with GM low DC or mature DC for 30,60 and 90 mins, then cpm value were determined. The changes in I(A)/I(E) and CD80 on cell surface after antigen ingestion were determined with flow cytometry (FCM). By using mixed lymphocyte reaction (MLR), the cells were divided into control (non-sensitized T lymphocyte), GM low DC, GM low DC and allogeneic antigen, GM low DC and allogeneic antigen and CTLA-4 Ig groups. The cpm value in each group was recorded and the stimulation index (SI) was calculated.</p><p><b>RESULTS</b>Upon 30, 60 and 90 mins of allogeneic Ag stimulation, the cpm value of GM low DC was obviously higher than that of mature DC (P < 0.05 or 0.01). In addition, the expression of I(A)/I(E) and CD80 before allogeneic Ag ingestion were significantly higher than those after Ag ingestion (I(A)/I(E): 32 +/- 8% vs. 54 +/- 10, P < 0.05; CD80: 25 +/- 10% vs. 71 +/- 18%, P < 0.01). MLR: Compared with control group, the cpm value in GM low DC with allogeneic Ag group was increased markedly (P < 0.05), with SI higher than 2.0, while no difference was found among control, GM low DC group, GM low DC and allogeneic Ag and CTLA-4Ig groups (P > 0.05), with SI lower than 2.0</p><p><b>CONCLUSION</b>Though GM low DC exhibits powerful antigen ingestion capacity, the cell phenotype and function will get mature gradually. Immune tolerance can be established by incubating GM low DC with CTLA-4Ig.</p>


Subject(s)
Animals , Female , Mice , Abatacept , Antigens , Allergy and Immunology , Dendritic Cells , Cell Biology , Allergy and Immunology , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , Immune Tolerance , Immunoconjugates , Pharmacology , Lymphocyte Activation , Mice, Inbred C57BL
12.
Chinese Journal of Burns ; (6): 423-426, 2006.
Article in Chinese | WPRIM | ID: wpr-331554

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the biological properties of immature dendritic cells( imDC) derived from cord blood before and after cryopreservation, so as to provide a method for preservation of imDC.</p><p><b>METHODS</b>Immature dendritic cells were generated from human cord blood (CB) monocytes and cultured with rhGM-CSF and rhIL-4, and 10% DMSO was added into culture medium as cryopreservation reagent. After freezing in - 80 degrees C refrigerator, the cells were finally cryopreserved in - 196 degrees C liquid nitrogen, and then thawed with 40 'C water, and they were finally named frozen-thawed imDC. The morphology of imDC were observed with light microscope, and TBR were calculated. Cellular surface markers for DC maturation were determined with flow cytometry, and the ability of the cells to stimulate proliferation of non-sensitized T lymphocyte was determined with allogeneic mixed lymphocyte reaction.</p><p><b>RESULTS</b>Monocyte (MNC) from cord blood could differentiate into DC after GM-CSF and rhIL-4 induction. Under light microscope, the cells showed irregular morphology, with branch-like prominence on the cell surface. Similar changes were also observed with scan electron microscope. The cryopreserved imDC were resistant to trypan blue staining, and TBR was (86. 8 +/- 1. 3) % . There was no obvious difference in the cell morphology between cryopreserved and fresh imDCs. The expression of cell surface markers and maturation markers in imDCs before cryopreservation were as follows: CDla(62 +/-8)% , CD14 (18 +/- 7)% , HLA-DR (67 +/- 5)% , CD80 (13+/-7)%, CD 86 (12+/- 5) % . Though the expression of CD80, CD86 and CD83 of cryopreserved imDC increased to (15 +/-5)% , (17 +/-5)% and (7.4 +/-3. 3)% , respectively( P <0.05), they still possessed the phenotype of imDC. There exhibited no obvious difference in cmp value between fresh imDC[ (463 +/- 104) min(-1) ] and cryopreserved imDC[ (512 +/-78 )min(-1) ] , ( P > 0. 05 ). The cpm in control group was (488 +/- 197 ) min'. The stimulation index in all groups was lower than 2, and both fresh imDC and cryopreserved imDC could not stimulate the proliferation of non-sensitized T lymphocyte.</p><p><b>CONCLUSION</b>The cryopreserved imDC exhibits immature characteristic in cell phenotypes, function and good cell activity, indicating that the method of cryopreservation of imDC is feasible.</p>


Subject(s)
Humans , Cell Differentiation , Cell Separation , Cryopreservation , Methods , Dendritic Cells , Cell Biology , Fetal Blood , Cell Biology , Flow Cytometry , Monocytes , Cell Biology
13.
Chinese Journal of Burns ; (6): 458-461, 2006.
Article in Chinese | WPRIM | ID: wpr-331544

ABSTRACT

<p><b>OBJECTIVE</b>To observe the changes in the phenotype characteristics and immune function after transfection of cord blood derived immature dendritic cells( imDC) with Adeasy-EGFP adenovirus vector, and to explore the function of IL-10 in inhibition of imDC maturation.</p><p><b>METHODS</b>Immature dendritic cells were generated from human cord blood(CB) monocyte cultured with rhGM-CSF and rhIL-4. The recombinant adenovirus vector AdEASY-EGFP was transduced into immature dendritic cells on the third day with or without adding IL-10. The expression of cell maturation marker CD83, CD86 and HLA-DR were determined with flow cytometry. Allogeneic mixed leukocyte reaction( MLR) was used to examine the imDC's ability to promote T cell proliferation.</p><p><b>RESULTS</b>The expression of surface maturation markers of imDC after transfection with adenovirus were significantly up-regulated ( CD86:46+/-10; CD83: 38 +/- 7; HLA-DR: 82 + 12) , and its ability to promote T cell proliferation was also obviously increased( SI > 2. 0). However, the expression of surface maturation markers of imDC after IL-10 treatment had lower mature phenotypes expression after transduction (CD86:8 +/- 5; CD83: 9 +/- 3; HLA-DR:63 +/- 12), and T cell stimulating ability was decreased comparing with adenovirus transfection groups.</p><p><b>CONCLUSION</b>Adenovirus can be transduced into imDC with high efficiency, but transfection itself can promote imDC's maturation. IL-10 treatment can inhibit the tendency to maturation stimulated by adenovirus transduction efficiently.</p>


Subject(s)
Humans , Adenoviridae , Genetics , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , Genetic Vectors , Interleukin-10 , Pharmacology , T-Lymphocytes , Metabolism , Transfection
14.
Chinese Journal of Surgery ; (12): 1219-1222, 2005.
Article in Chinese | WPRIM | ID: wpr-306132

ABSTRACT

<p><b>OBJECTIVE</b>To study the dynamic effects of allogenic transplantations with the bladder acellular matrix grafts (BAMG) of rabbits.</p><p><b>METHODS</b>Hemi-cystectomies were performed in 25 rabbits, and the defects were repaired with BAMG about half bladder size. The rabbits underwent postoperative assessment of bladder function at 8 weeks, including cystometry, vesical volume, vesical compliance and cystography. The allografts were observed by light microscope and electron microscope at 1, 2, 4, 8, 12, 16 weeks after surgery.</p><p><b>RESULTS</b>Macroscopic observation revealed that BAMG regenerated gradually. All urodynamic results of 8 weeks after surgery were not different statistically as compared with these of preoperation (P > 0.05). Cystography revealed that the morphous of bladder was recovered. Epithelialization and neovascularity occurred accompanied by infiltration of inflammatory cell at 1 week. Smooth muscle cell and stratified epithelium regenerated 2 weeks after grafting. Neural elements formed around smooth muscle bundles as early as 4 weeks. Each component regenerated on the frame of BAMG sequentially. After 16 weeks, it was difficult to delineate the junction between the host bladder and BAMG by histology.</p><p><b>CONCLUSION</b>After allogenic transplantation with rabbits' BAMG, the constitution and function of the allografts regenerate completely and gradually on the frame of BAMG.</p>


Subject(s)
Animals , Rabbits , Extracellular Matrix , Transplantation , Regeneration , Tissue Engineering , Transplantation, Homologous , Urinary Bladder , Cell Biology , Physiology , Transplantation
15.
Chinese Journal of Biotechnology ; (12): 466-472, 2005.
Article in Chinese | WPRIM | ID: wpr-305249

ABSTRACT

Superoxide dismutase, catalase and hemoglobin were purified simultaneously from the same batch of bovine erythrocyte lysate. The process involves an initial anion exchange chromatography, followed by a hydrophobic interaction chromatography and gel filtration chromatography. 0.75% polyethylene glycol 600 was added as a purification chaperon before the anion exchange chromatography. The hemoglobin fraction passed through the ion exchange column without being retained. The superoxide dismutase and catalase were adsorbed by the column and were eluted separately during elution. The two eluted fractions containing crude superboxide dismutase and catalase were further purified with hydrophobic interaction chromatography and gel filtration chromatography in sequence. The specific activities of superoxide dismutase and catalase were 15932u/mg and 65918u/mg, respectively. SDS-polyacrylamide gel electrophoresis and gel filtration chromatography were used to analyze the purity of the proteins. The purity of superoxide dismutase, catalase and hemoglobin were 77.6%, 81.9% and 99.9%, respectively. The total recoveries for superoxide dismutase, catalase and hemoglobin were 47.4%, 29.6% and 88.7%, respectively.


Subject(s)
Animals , Cattle , Catalase , Chromatography, Ion Exchange , Erythrocytes , Chemistry , Glucosides , Chemistry , Hemoglobins , Polyethylene Glycols , Chemistry , Superoxide Dismutase
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