Objective To evaluate the utility of fluorescent dye SYTO 13 for high -resolution melting ( HRM) detection in single nucleotide polymorphism ( SNP) genotyping and its clinical application . Methods This is a performance verification study .36 genotype defined samples were divided into three groups:SNP rs3125734 C>T (class Ⅰ SNP) ,rs255758 A>C (class ⅡSNP) and rs688C>T.These samples were used to evaluate SYTO 13′s SNP genotyping capability of class ⅠSNP, classⅡSNP, and two PCR products of different lengths (52 and 107 bp) covering the same SNP of rs688C>T.The commercial HRM dye of LCGreen Plus was used as the control .The genotyping capability is indicated by the Tm difference(ΔTm) between wild type and homozygous mutant genotypes .The Tm differences between wild genotype and homozygous mutant genotype were compared using the Independent Samples t test.Paired t test was used to evaluate genotyping capability of the two dyes .The clinical applicability is evaluated by synchronously performing PCR amplification and HRM analysis on thirty -five randomly selected DNA samples with known genotypes of the three SNPs .Results The SNPs of class Ⅰ and class Ⅱ can be genotyped directly and clearly with SYTO13 (ΔTmclas Ⅰ =0.36 ±0.05,tclas Ⅰ =14.827,Pclas Ⅰ =0.000;ΔTm clas Ⅱ =0.42 ±0.110,tclasⅡ =9.539,Pclas Ⅱ =0.000).The classⅠSNP genotyping results was better using SYTO13 (ΔTmSYTO13 =0.39 ±0.027), while the SNP genotyping for small amplicon did not discriminated clearly in this study .Long amplicons of class ⅠandⅡSNPs can be identified directly except for several samples which can be genotyped accurately after having performed reexamination .Conclusion SYTO13 can apply for HRM analysis of genotyping classⅠand ⅡSNPs with long amplicon and for clinical routine detection.

Bilingual teaching is adapted to the development of higher education in china.Based on actual fact of college,teaching mode,evaluation and effect of bilingual teaching on medical microbiology were studied,which started with necessity of bilingual teaching to use original edition teaching material in English. The result would provide some gist to choice the suitable pattern of bilingual teaching for other subject of our college.

Objective To study the clinical feature,treatment,and prognosis of the cytomegalovirus (CMV)pneumonia patients treated with immunosuppressor against kidney disease.Mlethod The patients received immunosuppressor against kidney disease in The First Affiliated Hospital of Sun Yat-sen University from June 1999 to December 2006.CMV antigen of leucocyte in the peripheral blood and/or bronchoalveolar lavage fluid of these patients were detected with immunocytochemical methods,and 21 patients were found suffering from CMV pneumonia.The 21 patients were introvenously injected with ganciclovir 5～10 mg/(kg?d),and the immunosuppressive agent treatment suspended.Their clinical feature and prognosis were retrospectively analyzed. Results The 21 patients received corticosteroids before CMV pneumonia contracted,of them,13 patients had been intensively treated with Methyllprednisolone with mean total dose(3.2?0.6)g.Of them,15 had been treated with cyclophosphamide with mean total dose(3.8?1.3)g.The median time from the beginning of using immunosuppressor to the onset of CMV pneumonia was 25(13～92)days.All patients had fever,cough, shortness of breath and X-ray showed interstitial pneumonia,of them,19 patients developed hypoxemia,and 11 patients' CMV antigen was positive in the leucocyte from bronchial lavage fluid.The result showed 9 patients survived and 12 died.The average duration of treatment with ganciclovir was(26.2?6.3)days. CMV pneumonia is a serious complication in patients who were treated with immunosuppressor against kidney disease.The mortality is high.Ganciclovir is a medicine of choice to treat CMV pneumonia.

Objective: To study the effect and mechanism of Shuanghua aerosol (SHA) in treating infantile upper respiratory tract infection. Methods: In the clinical study, 276 patients were randomly divided into two groups. The treated group was treated with SHA and the control group was treated with Shuanghuanglian aerosol (SHLA, 双黄连气雾剂). In the experimental study, the effects of SHA on anti-inflammation and anti-virus were observed. Results: The clinical total effective rate of SHA was 99.03% and its cure rate 65.38%, while those of the SHLA were 94.11% and 44.12% respectively, significant difference was shown between the two groups (P<0.01). Experimental study showed that SHA could inhibit the xylol induced ear swelling and the egg white induced paw swelling in mice obviously, and inhibit the proliferation of influenza virus in rat's lung. Conclusion: SHA has obvious anti-inflammatory and anti-viral effect, and has good curative effect in treating infantile upper respiratory tract infection.

Objective To evaluate the safety of 3D printed titanium alloy trabecular reconstruction rod, bone grafts and its clinical efficacy. Methods Sixty patients with osteonecrosis of femoral head (ONFH) stageⅡ(ACROⅡ) were randomly divided into experimental group (titanium rod group) and control group (tantalum rod group). Patients were followed up at 6, 12 and 24 months after the operation. Harris score and visual analogue scale (VAS) were used to evaluate the postoperative function. Changes of X-ray were observed before and after operation. The imaging evaluation was analyzed after 24 months. Results The Harris score was decreased in titanium rod group compared with that of tantalum rod group at six-month. There were no significant differences in Harris scores for other time points between two groups. Repeated measures analysis of variance showed that there was a significant difference in time effect. The X-ray staging results changed gradually before and after operation in two groups, stage Ⅲand stage Ⅳgradually increased with time. There was progress in ONFH after operation in two groups. No infection or rejection was found in the two groups. There were higher improvement rates of stageⅡA and stageⅡB. There were no significant differences in improvement rate and hip preservation rate between two groups. The transparent lines disappeared in the positive and lateral X-ray films 6 months after the operation. No new transparent lines were found. The bone enhancement was not found in two groups of patients during follow-up period. No stress occlusion and bone trabecular bone enhancement were observed in two groups. Conclusion Titanium rods are satisfactory for the treatment of femoral head necrosis in the early stage of ACRO Ⅱ, which is not recommended for the treatment of relatively large ACROⅡC stage of necrotic area.

Objective To investigate the value and dynamic characteristics of diffusion-weighted imaging(DWI)in rabbit liver VX_2 tumor model,with correlation of pathology.Methods Forty New Zealand rabbits were included in the study and VX_2 tumor piece was implanted directly into the liver after laporotomy.Fiffteen had two intrahepatic implants while twenty-five had one implant.DWI was performed on the seventh,fourteenth and twenty-first day after implantation,while routine T_1WI and T_2WI sequences were done on the seventh and fourteenth day.Ten VX_2 tumor samples were studied by pathology.Results The lump detection rates on the seventh day after implantation of DWI,T_1WI and T_2WI were 78.7%(37/47), 10.7%(3/28)and 53.6%(15/28)respectively with statistical significance(x~2=32.61,P

Objective To observe the effect of berberine on visfatin expression in 3T3-L1 adipocytes,and to explore the mechanism underlying berberine improving insulin resistance.Methods Visfatin mRNA and protein expressions were assayed by RT-PCR and Western blot after incubation of berberine at different concentrations and durations with 3T3-L1 adipocytes.Results In 3T3-L1 adipocytes,visfatin mRNA expression was increased at 0- 10?mol/L berberine with the maximum effect at 10?mol/L,being 4.96 folds of control group.While as the berberine concentrations were further increased above 10?mol/L,the visfatin mRNA level decreased.After incubation with berberine for 3 h,the visfatin mRNA level began to increase,and reached the peak when incubation for 12 h(4.57 fold of basal group).Then the mRNA level decreased,though it was still higher than that in control group after 48h intervention.5,10, 20?moL/L berberine could increase visfatin protein expression by 1.13,2.46,2.34 folds respectively as compared with control group(P

Mouse L1210 leukemia cell line is widely used as a model in the study of tumorigenesis, as well as the efficacy of chemotherapeutic drugs; however, like other suspension cell lines, the mouse L1210 cell line has lowest transfection efficiency, that many barriers exist to study about the structure, function, as well as metabolism in leukemia cells. This study was aimed to obtain higher transfection efficiency of L1210 cell line to facilitate scientific research. The transfection efficiencies of nucleofector and liposome in L1210 leukemia cells were detected by converted fluorescence microscopy and flow cytometry using EGFP (enhance green fluorescent protein); cell viability was observed by trypan blue exclusion test. The results showed that the transfection efficiency of nucleofector primarily through reporter gene pEGFP by Amaxa Nucleofector(TM) nuclear transfer apparatus was significantly higher than lipofectamine 2000 transfection, furthermore, in the same cell density (2 × 10(6)/ml) and plasmid content (10 µg), the transfection efficiency of nuclear transfer apparatus default mode A-20 was higher than that of other modes (S-18, T-20). Its survival rate was up to 50.5% after 24 hours. Cell viability of liposome transfection reached to 88% after 24 hours, but the transfection efficiency was lower (< 1%). It is concluded that the nuclear transfer apparatus A-20 transfected L1210 can reach higher transfection efficiency up to 61.6%, which is significantly higher than that of lipofectamine transfection. The survival rate is up to 50.5% well meeting the needs of scientific research. Higher transfection efficiency is helpful for in-depth research about the morphology, functions and pathogenesis in leukemia model L1210, and provides more searching space for the treatment of leukemia diseases.
Animals ; Cell Line, Tumor ; Cell Nucleus ; genetics ; Cell Survival ; genetics ; Genes, Reporter ; Green Fluorescent Proteins ; genetics ; Liposomes ; Mice ; Transfection ; methods

OBJECTIVETo analyze occurrence, prevention and treatment of the complications of thoracoscopic assisted spine surgery.

METHODSRetrospective review of 182 patients who underwent standard thoracoscopic technique or video-assisted thoracic surgical procedure from October 1998 to August 2004. The treatment of thoracic diseases included debridement, decompression with (or) reconstruction. The total number of complications were recorded, and its mechanism, prevention and treatment were analyzed.

RESULTSComplications occurred in 16 patients, 12 cases of perioperative complications included 3 patients suffered from pneumonia, 3 pulmonary atelectasis, 2 patients' lung injured by trocar, 1 patient obtained transient monoplegia, 2 suffered from transient intercostal nerve pain and 1 had superficial incision infection. Long-term complications occurred in 4 cases: spinal tuberculosis relapsed 2 cases (one who had diabetes obtained relapse in 8 months of post-operation and another relapsed with complex spinal tuberculosis in 4 weeks postoperation), 2 patients suffered from kyphosis deformity and pain.

CONCLUSIONSThe type and incidence of complications with thoracoscopic spine surgery mainly depend on indication, operation procedures and anesthesia, only by limit surgical indication, ameliorate technique, obey surgical principle and consummate perioperative treatment can we obtain mini-invasive effect by thoracoscopic assisted spine surgery.

Adolescent ; Adult ; Decompression, Surgical ; adverse effects ; methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spinal Diseases ; surgery ; Spinal Fusion ; adverse effects ; methods ; Thoracic Vertebrae ; surgery ; Thoracoscopy ; adverse effects

OBJECTIVETo study the early X-ray and CT findings of patients with severe acute respiratory syndrome (SARS).

METHODSChest radiography and CT were performed in 28 patients with SARS within one to three days after onset of the disease. CT examinations included conventional spiral CT and high-resolution CT (HRCT). The radiographic and CT findings of these patients were analyzed retrospectively.

RESULTSAbnormal CT findings were noted in all the patients, but abnormal chest radiographic findings in 17 cases (60.7%, 17/28). CT showed single small focal patchy opacities in 23 patients (82.1%, 23/28), including oval ground-glass opacities in 20 patients, lobular distribution ground-glass opacities in 2 and small patchy consolidation in one. Multi-focal ground-glass opacities were found in 2 patients and extensive opacities in three. In the 28 patients, a total of 31 lesions were found in the upper (7, 22.6%), middle (3, 9.7%), and lower lobes (21, 67.7%). The diameter of the lesions ranged from 20 to 35 mm.

CONCLUSIONThe dominant feature of early SARS patients is focal patchy opacity in the lung, and oval small ground-glass opacities are the common morphological findings on CT.

Adult ; Female ; Humans ; Male ; Middle Aged ; Radiography, Thoracic ; Severe Acute Respiratory Syndrome ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed