Animals ; Genes, BRCA2 ; Genes, erbB-1 ; Genes, erbB-2 ; Genes, p16 ; Genes, p53 ; Genes, ras ; Genetic Predisposition to Disease ; genetics ; Humans ; Pancreatic Neoplasms ; classification ; genetics ; pathology

Objective To explore a method for isolation and culture of human epidermal stem cells. Methods Epidermis was obtained by digesting human foreskin with Dispase Ⅱ and Trypsin-EDTA.After suspension on the epidermal stem cell medium (ESCM), these single epidermis cells were inoculated onto human collagen Ⅳ-coated flasks and cultured at 37 ℃ in a humidified atmosphere containing 5% CO_2 for 10 min. The nonadherent cells were rinsed off 10 min after inoculation, and the adherent cells continued to be cultured after enriching and abstraction by type Ⅳ collagen. The cell growth was observed through inverted microscope, and the cell cloning efficiency and time of clone sustain were also detected. Immunocytochemistry was used to observe the expression of ?_1-integrin and keratin 19(K19). Keratinocytes were served as controls. Results It was revealed by histological observation that colonies were formed 24 hours after inoculation. The isolated and cultured cell cloning efficiency was higher and the time of clone sustain was longer than that of the control group. Positive expression of ?_1-integrin and K19 of cultured cells was detected by immunocytochemistry. Conclusion Adult epidermal stem cells could be successfully isolated and cultured by adhension with type Ⅳ collagen and culture with ESCM.

OBJECTIVETo investigate the expression of TGF-beta1 mRNA in prostate carcinoma tissues and its significance.

METHODSRT-PCR method was used to examine the expression of TGF-beta1 mRNA in normal and carcinomatous prostate tissues.

RESULTSThe relative content of TGF-beta1 mRNA of normal prostate tissues was (0.74 +/- 0.11), while those of carcinomatous prostate tissues at T1-T2 and T3-T4 stages were (0.69 +/- 0.10) and (0.44 +/- 0.08) respectively, with significant difference between T1-T2 and T3-T4 (P < 0.05). And the relative contents of TGF-beta1 mRNA of carcinomatous prostate tissues with Gleason score < or = 5, 6-8 and > or = 9 were (0.70 +/- 0.12), (0.54 +/- 0.11) and (0.42 +/- 0.09) respectively.

CONCLUSIONThe expression of TGF-beta1 mRNA was negatively correlated with the clinical stage and Gleason score of prostate carcinoma.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; biosynthesis ; genetics

Objective To evaluate the clinical features and the strategy for the diagnosis and treat- ment of bilateral testicular tumor.Methods The clinical data (including the signs and symptoms,imaging studies,tumor markers,treatment modalities and histopatbologic diagnoses) of 10 cases of bilateral testicular tumor from January 1980 to December 2004 were reviewed.Their age ranged from 19 to 58 years(mean,34 years).Of the 10 cases,8 with metachronous and 2 with synchronous testicular tumors were identified.The clinical stages at the primary and secondary tumor diagnosis were:5 cases of stageⅠ,3 of stageⅡ;and 6 cases of stageⅠ,1 of stageⅡ,and 1 of stageⅢ,respectively,in 8 metachronous tumor patients.Two syn- chronous tumor patients were both identified as stageⅠdisease.Histological examination showed the primary tumor (seminoma) in 4 cases and the secondary contralateral tumor (seminoma) in 3.Results Two syn- chronous tumor patients underwent bilateral radical orchiectomy simultaneously,and 8 underwent orchiectomy successively.Retroperitoneal lymph node dissection was performed in 3 cases.Postoperatively,hypogonadism occurred in 10 patients,and 7 of them received androgen replacement therapy.Follow-up ranged from 9 month to 23 years with a mean of 10.5 years.Two patients died of the disease;2 had metastasis (1 of them was alive with metastasis);2 had recurrences and underwent local resection.Conclusions Metachronous bilateral testicular cancers are more common than synchronous bilateral testicular cancers.Seminoma was the most common histopathologic type.Testis-sparing surgery can be performed in selected cases.

Objective To investigate the reversal effect of MDR1 and MDR3 gene silencing on resistance of A2780/taxol cells to paclitaxel.Methods shRNA plasmid vector specifically targeting MDR1 and MDR3 genes was transfected into A2780/taxol cells.The early stage cell apoptosis and the effect of intracellular rhodamine 123(Rh123)accumulation were detected by flow cytometry(FCM).The late stage cell apoptosis rate was detected by terminal deoxynucleotidyl transferase(TdT)-mediated deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL).The 50% inhibition concentration(IC_(50))of paclitaxel on A2780/taxol cells was determined by methyl thiazolyl tetrazolium(MTT)assay.MDR1 and MDR3 mRNA were assessed by RT-PCR,and caspase-3 protein was detected by western blot.Results After treatment with MDR1 and MDR3 shRNA plasmid vector,early apoptosis rate of A2780/taxol cells was (20.21?0.56)% and(10.87?1.24)%,respectively.MDR1 and MDR3 shRNA could increase cellular Rh123 accumulation(116.6?8.1 and 98.4?3.8,respectively).The late stage apoptosis rates detected by TUNEL displayed the same tendency as FCM results did.The IC_(50)for paclitaxel of A2780/taxol cells was decreased significantly.The mRNA levels of MDR1 and MDR3 in A2780/taxol cells were decreased by (73.3?0.8)% and(51.6?0.4)% of control,and the reduction of MDR1 and MDR3 mRNA was in a time-dependent manner.The expression of caspase-3 protein of MDR1 and MDR3 shRNA vector transfected group in A2780/taxol cells was significantly increased [(80.8?2.6)% and(72.0?4.7)%, respectively ].Conclusion MDR1 and MDR3 gene silencing could recover sensitivity of A2780/taxol cells to paclitaxel and induce cell apoptosis,thus reversing cell resistance to paclitaxel.

OBJECTIVETo investigate the experience on diagnosis and treatment of multiple adrenal aldosterone-producing adenomas (APA).

METHODSEighteen cases of multiple adrenal APA were analyzed retrospectively, which were admitted from October 1992 to April 2006.

RESULTSAdrenalectomy was performed for 4 cases of unilateral synchronous multiple APA, which were discovered with three adenomas by 3D-CT; bilateral tumor resection was performed for 6 cases of bilateral synchronous multiple APA. There were 8 cases of bilateral metachronous multiple APA, including 2 cases of ipsilateral recurrent adrenal APA after adrenal tumor removal, which underwent tumor resection. Another 6 cases were contralateral APA following adrenalectomy due to adrenal APA, and underwent tumor resection. After operation, the adrenal function seemed to be normal, and no recurrence had been found on follow-up.

CONCLUSIONSUnilateral multiple synchronous APA require adrenalectomy. Tumor resection should be performed for bilateral or asynchronous APA, and it is very important to preserve healthy adrenal tissue as much as possible. 3D-CT has much value on diagnosis of small APA, unilateral multiple synchronous APA and ipsilateral recurrent adrenal APA.

Adenoma ; complications ; diagnosis ; surgery ; Adrenal Gland Neoplasms ; complications ; diagnosis ; surgery ; Adrenalectomy ; Adult ; Aldosterone ; blood ; Female ; Follow-Up Studies ; Humans ; Hyperaldosteronism ; blood ; etiology ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo observe the effect of Kanglaite injection(KLT) on the proliferation and telomerase activity of mesangial cells in rats.

METHODMTT, telomere repeat amplification protocal (TRAP), ELISA, PAGE and silver-stain were applied to detect the growth rate and telomerase activity of MC after stimulation of KLT and IL-1.

RESULTThe growth rate of MC was enhanced by IL-1 stimulation, which was accompanied with a redection of the activity of telomerase. Adversely, the growth rate of MC was reduced by KLT, which was accompanied with an enhancement of activity of telomerase. Moreover, the growth rate of MC and the activity of telomerase were both inhibited by the combinative use of IL-1 and KLT without any influence from the sequence of their administration.

CONCLUSIONKLT could inhibit proliferation and telomerase activity of MC with or without pre-stimulation with IL-1. KLT might be useful to prevent and treat glomerular nephritis related to MC proliferation.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Coix ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Glomerular Mesangium ; cytology ; enzymology ; Injections ; Plant Oils ; administration & dosage ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rats ; Seeds ; chemistry ; Telomerase ; metabolism

OBJECTIVETo investigate the effect of recombinant human interleukin-2 (rhIL-2) activated natural killer cells (rhIL-2-NK) on angiogenesis and cardiac function of rats with myocardial infarction (MI).

METHODSNatural killer cells (NKs) were isolated and activated by rhIL-2 in vitro. Untreated NKs were used as the control, the killing capacity of rhIL-2-NK were evaluated with cytotoxicity assay. Cardiac microvascular endothelial cells (CMECs) were cocultured with rhIL-2-NK. One hour after MI, rats were randomly divided into rhIL-2-NK group, NK group and blank control group and NK, rhIL-2-NK and PBS were injected directly in the infracted myocardium. At the 0, 1(st), 3(rd), 5(th), 7(th) and 20(th)th day after MI, the mRNA expression of monocyte chemotactic protein-1 (MCP-1), Tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) were was detected by q-PCR essay. At the end of the therapy, the platelet endothelial cell adhesion molecule-1(CD31) and vascular endothelial growth factor (VEGF) were evaluated through immunohistochemical assay, and the cardiac function observed with echocardiography, homodynamic measurements.

RESULTSThe NKs were isolated successfully and the CMEC were proliferated remarkably by coculturing with rhIL-2-NK (P < 0.01). The mRNA expression of MCP-1, TNF-α, CD31 and rhIL-2, VEGF were significantly upregulated in rhIL-2-NK group than in the PBS control group (P < 0.01). Four weeks after operation, LVEF was significantly higher in rhIL-2-NK group than in the PBS control group [(77.56 ± 15.67)% vs. (41.47 ± 12.21)%, P < 0.05)] and histomorphology assay revealed that the density of microvascular endothelial (MVD) of rhIL-2-NK group was significantly higher than that of PBS control group (17.35 ± 1.82 vs. 4.76 ± 0.92, P < 0.01).

CONCLUSIONSMyocardial injection of rhIL-2-NK could promote angiogenesis and improve cardiac function in MI rats.

Animals ; Heart ; physiology ; Humans ; Interleukin-2 ; pharmacology ; Killer Cells, Natural ; drug effects ; physiology ; Myocardial Infarction ; physiopathology ; Neovascularization, Physiologic ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacology

Objective To explore the associations between vitamin D receptor ( VDR) Fok- Ⅰ and Apa- Ⅰ polymorphisms and myasthenia gravis (MG) in Chinese Han population.Methods Polymorphisms of VDR Fok- Ⅰ and Apa-Ⅰ were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.The frequencies of genotypes and hyplotypes were compared among 286 normal controls and 267 MG patients in different subgroups classified by gender,age of onset,presence of thymoma,and Osserman classification sat maximal severity in the follow-up.The association between the genotypes and maximal severity of MG and short-term glucocorticoid treatment were also investigated.Results There were no significant differences in frequencies of genotypes and hyplotypes of both Fok-Ⅰ and Apa-Ⅰ between MG group and control group,and among subgroups of MG.The Fok- Ⅰ showed no statistical difference between the patients with better and less improvement after short-term treatment of glucocorticoid.The frequency of Apa-Ⅰ alleles in the patients with better improvement (55/186,29.6% ) significantly differed from the less improved group ( 7/48,14.6%,OR = 2.46,95% CI 1.04-10.43,x2 = 4.400,P = 0.036).The patients with the genotype A A/Aa were more likely to improve better after the treatment(48/93,51.6%) than in the worse group(7/24,29.2%,OR =2.59,95% CI 0.98-14.60,x2 =3.858,P= 0.049).Conclusions Alleles and genotypes of VDR-Fok- Ⅰ and Apa-Ⅰ were not found to be related with MG onset and severity.MG patients with VDR-Apa-A allele may have better improvement short-term treatment of glucocorticoid.

OBJECTIVETo establish a stable high red fluorescent protein (RFP)-expressing orthotopic transplantation nude mice spontaneous metastasis model of pancreatic cancer.

METHODSStable high RFP-expressing cells SW1990-RFP were injected subcutaneously into mice to establish subcutaneous implantation model. Fluorescent tumor piece from subcutaneous was transplanted into the body of the pancreas to establish surgical orthotopic implantation model. The growth of primary tumor, metastasis and micrometastasis were assessed by whole-body fluorescence imaging system.

RESULTSTwelve RFP orthotopic transplantation nude mice metastasis models of pancreatic cancer were established successfully, the percentage of success rate was 100%. RFP-labeled pancreatic cancer growth could be monitored in real time way. The micrometastasis of primary lesions were detected in early stage with whole-body fluorescence imaging system.

CONCLUSIONSThe RFP orthotopic transplantation nude mice metastasis model of pancreatic cancer is stable and reliable, and can be observed dynamically in vitro in a noninvasive way, with much higher sensitivity and specificity.

Animals ; Disease Models, Animal ; Female ; Luminescent Proteins ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Pancreas ; pathology ; Pancreatic Neoplasms ; pathology ; Xenograft Model Antitumor Assays