To obtain high-yield avilamycin-producing strains,low energy N~+ ion implantation technology and screening of streptomycin-re- sistant mutants are used in the study on breeding mutation.The results show that,“saddle”region,which range is from 3?10~(15) to 5?10~(15) ions/cm~2,has got better induced mutation action.It also means that the strain's resistant mutation and yield mutation closely correlate to each other,and the method of streptomycin resistant screening is feasible.We have isolated a high-yield strain SVT-45 which the productivi- ty is 195% higher than the original strain's in the rotation-flask experiments.These results showed that the ion implantation was an effective method for microbe mutagensis.

Background The rapid diagnosis can win more treating opportunities for patients with fungal keratitis.Even though the fungal culture is the gold standard for the diagnosis of fungal keratitis,it is difficult in early diagnosis due to the long duration of cultivation and false-negative rate.Objective This trial was to explore the clinical value in the rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy.Methods Corneal scraping and laser scanning confocal microscopy were separately performed in 167 eyes of 167 patients with fungal keratitis.All the eyes were examined by the slit lamp,followed by laser scanning confocal microscope,and then the 10％ KOH corneal smear was examined under the optical microscope.Results The positive rate of diagnosis was 75％ (125/167) by corneal scraping,and that by laser scanning confocal microscopy was 91％ (152/167).The positive rate of examining outcome was significantly higher in laser scanning confocal microscopy than that of corneal scraping (x2 =14.88,P =0.00).The positive results were 114 cases and negative results were 4 cases by two methods,with the concordance rate 70.7％ (118/167).The hyphae or spore were seen in 32 cases by laser scanning confocal microscopy in 42 negative cases by corneal scraping,and in 15 negative cases by confocal laser scanning microscopy,11 positive outcomes were offered by corneal scraping.Conclusions The combined application of corneal scraping with confocal laser scanning microscopy can improve and speed up the diagnosis positive rate of fungal keratitis.

Objective To investigate the changes of iodide uptake and the expression of thyroidspecific genes in poorly differentiated follicular thyroid carcinoma (FTC) cells after transfection of human TSH receptor (hTSHR) gene in vitro. Methods The recombinant eukaryotic expression plasmid PcDNA3. 1/hTSHR-cDNA was transformed into DH5a bacterial for amplification and then the recombinant plasmid was extracted. The recombinant was identified with PCR amplifying, restriction enzyme digestion analysis and DNA sequencing. The recombinant plasmid pcDNA3.1/hTSHR was transfected into FTC-133 cell line by lipofectin methodin vitro. Immunofluorescence, iodide uptake studies and real time-PCR were applied to detect target protein expression. Statistical analysis was performed with t-test using SPSS 13. 0 software. Results Kpn Ⅰ and Xba Ⅰ restriction enzyme digestion, PCR amplifying and DNA sequencing confirmed that pcDNA3. 1/hTSHR was successfully constructed. After transfection of the recombinant plasmid pcDNA3. 1/hTSHR-cDNA and the stimulation of hTSH, the tumor cells displayed the expression of hTSHR protein at cell surface and cytoplasm. The iodine uptake in pcDNA3. 1/hTSHR transfected cells was 2. 9 times higher than that of control(pcDNA3.1(+) transfected cells) group(t = 28.63, P ＜0. 01). The expression of TSHR,NIS, TPO and Tg (mRNA levels) in pcDNA3. 1/hTSHR transfected cells were also significantly elevated by 1.74 (t =5.959, P＜0.01), 7.2 (t =3.807,P＜0.05), 2.88 (t=4.769,P＜0. 01) and 2.67 times (t=6.388,P ＜0.01) respectively compared to those of the control group. Conclusion The study demonstrates that iodide uptake may be reactivated by hTSHR receptor gene transfection in poorly differentiated FTC cell.

OBJECTIVETo investigate the association between transforming growth factor beta-1 (TGF-beta1) gene polymorphism and chronic allograft nephropathy (CAN).

METHODSFifty patients with failed renal allografts and clinically and histopathologically confirmed CAN were enrolled in this study along with another 50 renal transplant recipients with normal graft function. The DNA extracted from whole blood of the patients was amplified with PCR with sequence-specific primers for determining TGF-beta1 genotypes (position +869, codon 10 and position +915, codon 25). According to documented descriptions, the patients were classified into high and moderate-to-low cytokine production genotypes. The distribution frequencies of high production genotypes was then compared between CAN and non-CAN groups. To eliminate interference in the analysis of the association between TGF-beta1 polymorphism and CAN, other possible risk factors for CAN were screened, including the patients' gender, age, HLA match, delayed graft function, acute rejection, immunosuppressive regimen, cytomegalovirus infection, hypertension, and high cholesterol.

RESULTSCAN patients showed significantly greater proportion of high cytokine production genotype than the non-CAN group [70% (35/50) vs 38% (19/50), Chi(2)=10.306, P=0.001). Of the screened risk factors for CAN, only acute rejection showed some difference between the two groups, but analysis after subgrouping according to acute rejection did not suggest its influence on CAN, which supports the result that the rate of high production genotype was significantly higher in CAN group than in the non-CAN group.

CONCLUSIONMost CAN patients have high TGF-beta1 production genotype, which might be a risk factor for CAN after renal transplantation. TGF-beta1 genotyping can be of value in predicting the risk of CAN after renal transplantation.

Adult ; Female ; Genetic Predisposition to Disease ; Graft Rejection ; genetics ; Humans ; Kidney Diseases ; genetics ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Polymorphism, Genetic ; Risk Factors ; Sequence Analysis, DNA ; Transforming Growth Factor beta1 ; genetics ; Transplantation, Homologous

OBJECTIVETo study the value of detection of both preoperative soluble CD30 (sCD30) and hepatocyte growth factor (HGF) level 5 days after transplantation in the diagnosis of acute rejection of renal allograft.

METHODSPreoperative serum sCD30 levels and HGF level 5 days after transplantation were determined in 65 renal-transplant recipients using enzyme-linked immunosorbent assay. The recipients were divided according to the sCD30 levels positivity. Receiver operating characteristic (ROC) curves were used to assess the value of HGF level on day 5 posttransplantation for diagnosis of acute renal allograft rejection, and the value of combined assay of the sCD30 and HGF levels was also estimated.

RESULTSAfter transplantation, 26 recipients developed graft rejection and 39 had uneventful recovery without rejection. With the cut-off value of sCD30 of 120 U/ml, the positivity rate of sCD30 was significantly higher in recipients with graft rejection than in those without (61.5% vs 17.9%, P<0.05). Recipients with acute rejection showed also significantly higher HGF levels on day 5 posttransplantation than those without rejection (P<0.05). ROC curve analysis indicated that HGF levels on day 5 posttransplantation was a good marker for diagnosis of acute renal allograft rejection, and at the cut-off value of 90 ug/L, the diagnostic sensitivity was 84.6% and specificity 76.9%. Evaluation of both the sCD30 and HGF levels significantly enhanced the diagnostic accuracy of acute graft rejection.

CONCLUSIONCombined assay of serum sCD30 and HGF levels offers a useful means for diagnosis of acute renal allograft rejection.

Enzyme-Linked Immunosorbent Assay ; Graft Rejection ; blood ; diagnosis ; Hepatocyte Growth Factor ; blood ; Humans ; Ki-1 Antigen ; blood ; Kidney Transplantation ; ROC Curve ; Sensitivity and Specificity ; Transplantation, Homologous

Objective To summarize the experience of long-term survival in patients after simulta- neous kidney-pancreas transplantation(SKPT)with modified enteric drainage(ED).Methods From October 2001 to July 2004,6 patients with end-stage renal disease due to Type 1 diabetes underwent SKPT with modified ED,ie,side-to-side anastomosis between the duodenum of donors and jejunum of recipients. The medication regimen included:mycophenolic acid 500 mg and tacrolimus 2 mg before operation;methyl- prednisolone(MP)1.0 during operation;and 2-dose anti-IL-2 receptor monoclonal antibody(2 cases)or antihuman thymocyte globulin(ATG)(4 cases)for immune induction therapy;MP was used on the first 3 d after transplantation,triple immunosuppressive therapy(tacrotimus,mycophenolic acid and prednisone)was used on the second d after transplantation.Anticoagulants such as low molecular heparin or alprostadil were used for 7-10 d to prevent thrombosis in pancreas graft.Somatostatin was used as prophylaxis for graft pan- creatitis.Ganciclovir was used to prevent cytomegalovirus infection when renal graft gradually recovered 3 to 5 d after transplantation.The follow-up was from 1 year and 3 months to 4 years and 1 month.Results Transplantation was successful in all 6 cases.The blood sugar levels were 6-16 mmol/L.Low-dose insulin was used for 5-10 d,then the blood sugar levels returned to normal range.One of 6 patients experienced nephrotoxicity because of high tacrolimus blood concentration at 7 d after operation;after 3 dialyses and re- duction of tacrolimus dose,the renal allograft regained normal function.Three cases experienced alimentary tract hemorrhage at 14,20 and 22 d,respectively,after operation;the bleeding was stopped after treatment. There were no complications such as pancreatic fistula,intestinal fistula and thrombosis early after operation. All the patients are now alive,specifically,1 survived over 4 years,3 over 3 years,1 over 2 years,and 1 over 1 year.All had normal blood sugar free of insulin use.Five cases had normal renal graft function,with normal sCr,and 1 had sCr＞400?mol/L. Two cases were admitted to hospital due to upper respiratory infection and furuncles in the skin of head 6 months and 2 years,respectively,after operation.They were both cured.No complications such as urinary infection,metabolic acidosis and dehydration occurred.Conclusions SKPT is effective for the treatment of end-stage renal disease due to Type 1 diabetes.SKPT with modified ED are relatively simple with physiological compatibility and fewer complications.High quality of donated organs, HLA matching,pancreatic drainage pattern,rational periopcrative medications and infection late after trans- plantation are important factors affecting the long-term survival of the patients.

OBJECTIVEAdrenocortical carcinoma (ACC) is a rare but highly malignant tumor, and its diagnosis is mostly delayed and prognosis is poor. We report estrogen receptor (ER) expression in this tumor and our clinical experiences with 17 ACC cases.

METHODSThe data of the 17 patients (9 females and 8 males, age range from 16 to 69 years, mean age of 42.6 years) with ACC were reviewed, and symptoms, diagnostic procedures, treatment, and results of follow-up were evaluated. Immunohistochemistry was used to detect ER expression in tumor samples from the 17 patients.

RESULTSAt the time of diagnosis, 4 tumors were classified as Stage I, 4 as Stage II, 3 as Stage III, and 6 as Stage IV. Eight patients demonstrated positive nuclear immunostaining of ER. The prognosis of patients with ER positive was significantly better (P<0.05) than that of patients with ER negative, with 1- and 5-year survival rates at 86% and 60% for ER-positive patients, and 38% and 0% for ER-negative patients, respectively.

CONCLUSIONER-positivity may be one of the factors associated with a worse prognosis of ACC.

Adolescent ; Adrenal Cortex Neoplasms ; diagnosis ; metabolism ; mortality ; Adrenocortical Carcinoma ; diagnosis ; metabolism ; mortality ; Adult ; Aged ; Biomarkers, Tumor ; analysis ; China ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; Receptors, Estrogen ; analysis ; Risk Assessment ; methods ; Risk Factors ; Survival Analysis ; Survival Rate ; Young Adult

Objective To evaluate the clinical value of multi-slice CT(MSCT)angiography in the diagnosis of Takayasu's arteritis.Methods Fourteen patients underwent MSCT angiography with use of 90 ml contrast media at a rate of 3.5 ml/s.Images were generated for vascular assessment with various reconstructional techniques,including multi-planar reformat(MPR),maximum intensity projection(MIP), volume rendering(VR),CT virtual endoscopy(CTVE)and advanced vessel analysis(AVA).Results MSCT angiography findings was consistent with the clinical changes of Takayasu's arteritis in all cases.The integrated information including reconstructional and transverse images clearly presented various luminal abnormalities,such as stenosis,occlusion and dilatation in the affected aorta,as well as its major branches and collateral circulation.Furthermore,it depicted mural abnormities,especially concentric arterial wall thickening.Steno-occlusive type(8/14)was more common than mixed type(6/14).According to Lupi- Herrea's classification,type Ⅰ in 5 cases,type Ⅱ in 2,type Ⅲ in 6 and type Ⅳ in 1 were recorded in the present study.Conclusion MSCT angiography is reliable imaging modality for the detection of Takayasu's arteritis.It is useful for early diagnosis because it allows evaluation of wall thickness rather than merely the luminal diameter,which is especially important for prognosis.It might be used as the first choice in the diagnosis of this disease as a simple,convenient and noninvasive method.

OBJECTIVETo study the changes in Notch1 expression on peripheral lymphocytes after acute graft rejection after renal transplantation.

METHODSTwenty renal transplant recipients experiencing acute graft rejection and 20 without acute rejection were enrolled in this study. Flow cytometry was used to detect the expression of Notch1 on peripheral lymphocytes of the patients before operation, at the occurrence of acute rejection and after anti-rejection therapy. The rates of Notch1-positive lymphocytes measured at different time points were compared between the two groups.

RESULTIn patients with acute graft rejection, Notch1 expression at the time of rejection onset was significantly higher than that before operation (t=4.245, P=0.000) and that of patients with graft rejection (t=3.839, P=0.000), and was obviously decreased after anti-rejection therapy (t=3.102, P=0.004). Patients without graft rejection showed no significant changes in Notch1 expression after the transplantation (P=0.409). Notch1 expression was comparable between the recipients receiving Tac therapy and those with CsA therapy (P>0.05).

CONCLUSIONMonitoring Notch1 expression on the peripheral lymphocytes after renal transplantation may help in the diagnosis of acute graft rejection and prediction of the effect of an anti-rejection therapy.

Adult ; Biomarkers ; blood ; Female ; Flow Cytometry ; Graft Rejection ; blood ; diagnosis ; Humans ; Kidney Transplantation ; adverse effects ; Lymphocytes ; metabolism ; Male ; Middle Aged ; Receptor, Notch1 ; blood

OBJECTIVETo investigate the genotypes of natural killer cell immunoglobulin-like receptor (KIR) genes and their frequencies in Chinese subjects and explore the mechanism of the actions of nature killer cells.

METHODSThe DNA samples were obtained from 67 randomly selected unrelated Chinese Han individuals for genotyping of the KIR genes using PCR with sequence-specific primers (PCR-SSP), and the frequencies of the KIR genes in these Chinese subjects were compared with the reported frequencies in populations of other nationalities.

RESULTSSixteen KIR genes were identified in these Chinese subjects, and 87.5% of these genes were expressed at frequencies above 0.35. Fourteen functional KIR genes combined into 25 KIR genotypes, among which the most frequent genotype KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 showed a frequency of 0.373, while the frequencies of all the other genotypes were no greater than 0.09. Comparison of the KIR combinations in Chinese Han population with those of Japanese, Korean, and Caucasians populations identified 8.93% of the KIR combinations shared by all these populations; the Chinese, Koreans and Caucasians shared 5.36% common KIR combinations, whereas only 1.79% common combinations were found in Chinese and Caucasians. In this study, 16 new gene combinations were identified (25.28%).

CONCLUSIONThis study shows the high-frequency distribution of a single KIR gene polymorphism. The KIR combination KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 has the highest frequency in Chinese, Japanese, Korean and Caucasian populations, indicating that inhibitory signal transduction pathway plays an important role in the function of the natural killer cells. This study provide clues for new approaches for improving the prognosis of kidney transplantation by enhancing or inhibiting the function of the natural killer cells instead of life-time usage of immunosuppressive agents.

Asian Continental Ancestry Group ; ethnology ; genetics ; Gene Frequency ; Genotype ; Humans ; Killer Cells, Natural ; immunology ; Polymorphism, Genetic ; Receptors, KIR ; genetics ; Sequence Analysis, DNA