Objective To assess the application value of REP-PCR in genotyping of candida glabrata strains in clinical pratice. MethodsFrom 2009 to 2010, thirty-eight candida glabrata strains were isolated from Shanghai Ruijin Hospitals, Shanghai Renji Hospital, Shanghai Huashan Hospital, Anhui Medical University Hospital, Shenzhen People's Hospital. Six loci in housekeeping genes (FKS, LEU2,NMT1, TRP1, UGP1 and URA3 ) were amplified and sequenced. The sequences were compared with the MIST database and allele profile and sequence type (ST) were obtained. With primers Ca21, Ca22 and Com21 used to amplify the adjacent variable gene regions,the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Finally, the results of these two genotyping methods were compared. ResultsFor REP-PCR, Ca22-Com21 has the best genotyping effect. REP-PCR and MLST have the same genotyping results. Five REP-PCR types were found in 38 candida glabrsta isolates. Type A,B, C, D and E strains from REP-PCR were genotyped as ST 7, 3, 19, 45 and new type respectively byMIST. REP-PCR saves time compared with MIST. Conclusions REP-PCR offers a simple and rapid method for molecular typing, with similar discriminatory power with MIST. Therefore, REP-PCR can be the preferred choice in laboratory, especially for a large number of isolates.

Objective To investigate four methods for establishing animal models of human breast cancer bone metastasis. Methods Thirty-two female nude mice aged 4-6 weeks were divided randomly into four groups (n=8 in each group). 5×105 MDA-MB-231 cells were injected into the body via the left second mammary fat pads (group A), the tail veins (group B), the left heart ventricles (group C) and the left tibia marrow cavities (group D), respectively. Tumor formations in situ were recorded in group A. Deaths after the injection were recorded. The surviving nude mice 49 days after the injection were subjected to pathological examination to determine bone metastasis. Results The rate of tumor formation in situ of group A was87.5 %(7/8). One mouse in group C died after the injection of MDA-MB-231 cells. The bone metastasis rate in groups A, B, C and D was zero (0/8), 12.5 % (1/8), 71.4 % (5/7) and 100 % (8/8), respectively. There was statistically significant difference in the bone metastasis rate between group A and group C, group A and group D, group B and group C; and group B and group D. Conclusion Injections of tumor cells via the breast fat pads and tail veins were not suitablemethods to establish animal models of human breast cancer bone metastasis. The bone metastasis model could be established efficiently by injecting tumor cells into the left heart ventricles or the bone marrow cavity of nude mice.

Objective To apply a fluorescent nucleotide dye Sybr Green I in detecting telomerase activity and assess its clinical value in the diagnosis of colorectal carcinoma. Methods Telomerase activity was (measured) by telomeric repeat amplification protocol (TRAP) combined with fluorescent nucleotide dye Sybr Green I in 46 cases of colorectal carcinoma tissue specimens and 19 tumor-adjacent tissue (specimens). Results (Telomeric) repeat amplification product was displayed as clear bands in PAGE stained with Sybr Green I; the positive rate of telomoerase activity in colorectal carcinoma tissue was 89.13%, whereas no telomerase (activity) was observed in 19 tumor-adjacent tissue specimens. There was no (relationship) between telomerase (activity) and the level of cell differentiation, Dukes stages, or metastasis of lymph nodes. Conclusions The method of TRAP combined with Sybr Green I to determine telemerase (activity) is an important method for the diagnosis of colorectal carcinoma.

Objective To investigate the utility of serum prostate specific antigen(PSA) density (PSAD),prostate antigen transition zone density(PSAT) and the ratio of free/total PSA with PSAD [(F/T)/PSAD] in the diagnosis of prostatic carcinoma (PCa) by three-dimensional ultrasonography.Methods Seventy-eight patients (serum prostate-specific antigen between 4-20 μg/L ) were involved.The prostatic volume and its transition zone volume were measured by three-dimensional ultrasonography.Then the relative parameters of PSA [PSAD,PSAT and (F/T)/PSAD] were calculated.Pathologic types were determined by using needle biopsy of prostate.Results Among them,27 patients were suffering from PCa,while the other 51 benign prostate hypertrophy (BPH).The difference of PSAD,PSAT and (F/T)/PSAD between PCa and BPH had arrived statistical significance (P<0.05).Proportions under the PCa curves were 0.736,0.760,0.800 respectively.Considering both sensitivity and specificity,a cutoff was recommanded:PSAD>0.20,PSAT>0.33,(F/T)/PSAD<0.8.Conclusions When the serum PSA level is between 4 μg/L and 20 μg/L,PSAD,PSAT and (F/T)/PSAD are of significant value to differentiate PCa from benign prostatic hyperplasia patients.The data are more reliable if prostatic volume are calculated by three-dimensional transrectal ultrasonography.

Objective To determine the effect of NEL-like type 1 gene (NELL-1) transfection in vivo in the repair of traumatic femoral head necrosis.Methods Twenty-four SD rats were randomly divided into three groups (8 rats per group) according to the lottery method,ie,sham group (served as normal control),NELL-1 treatment group (injected NELL-1 gene by recombinant adenovirus vectors around the hip one week after osteonecrosis model induced surgically) and placebo group (given an equal volume of saline solution at the same time after the induction of osteonecrosis).Femurs were taken from the animals 5 weeks after surgery.Gross observation was performed for morphology changes,X-ray assessment for femoral head height and length ratio (H/L),Micro-CT measure for bone parameters of femoral head including total volume (TV),bone volume (BV),total mineralized content (TMC),trabecular thickness (Tb.Th) and trabecular space (Tb.SP),and histological study for osteocytes,osteoblasts and osteoclasts.Results Preserved femoral head shape was noted in NELL-1 treatment group compared to the obvious flattening of the femoral head in placebo group.No heterotopic osteogenesis was observed in any group.Femoral head H/L ratio for 0.753 2 ± 0.040 2 in NELL-1 treatment group was higher than 0.598 4 ± 0.037 0 in placebo group (P ＜ 0.05),but lower than 0.920 2 ± 0.037 0 in sham group (P＜0.05).TV,BV,TMC and BMD between NELL-1 treatment and sham groups did not differ significantly (P ＞ 0.05),but all were increased compared to placebo group (P ＜ 0.05).There was no significant differences in Tb.Th and Tb.SP among three groups (P ＞ 0.05).Most osteocytes were alive in NELL-1 treatment group.More active osteoblasts and osteoclasts were noted in NELL-1 treatment group than those in placebo group.Conclusion NELL-1 gene transfection can preserve femoral head shape and bone content,promote osteoblast activity and neovascularization and hence is an effective treatment for rat traumatic osteonecrosis.However,the activity of osteoclasts is stimulated simultaneously.

OBJECTIVETo observe the efficacy and safety of Qizhi Jiangtang Capsule (QJC) in treating stage 3b diabetic kidney disease (DKD) patients with macroalbuminuria.

METHODSPatients who conformed to the diagnostic criteria of stage 3b DKD were randomly assigned to two groups according to random digital table, the experiment group and the control group, 84 in each group. All patients received a two-week elution period, and then were treated with basic Western therapy. Patients in the experiment group took QJC, 5 pills per time, 3 times a day, while those in the control group took Valsartan Capsule 160 mg each time, once daily. The observation period of follow-ups was limited within 6 months, and the time points were set as the baseline, 1st month, 3rd month, and 6th month. Systolic blood pressure (SBP), diastolic blood pressure (DBS), 24 h urine protein quantitative (24 h UPQ), plasma albumin (ALB), and serum creatinine (SCr) were detected and recorded, and estimated glomerular filtration rate (eGFR) was calculated. The occurrence of hypoglycemic reaction, coagulation disorder, gastrointestinal tract reaction, allergy, hyperkalemia, doubling of creatinine, and overall adverse events were observed and recorded at same time.

RESULTSFinally 81 patients in the experiment group and 80 patients in the control group were effectively included. Compared with the baseline level, SBP and DBS obviously decreased in the control group at month 1 of treatment (P < 0.05), and more significantly decreased at month 6 of treatment (P < 0.01). SBP at month 1, 3, and 6 of follow-ups; DBS at month 6 of follow-ups was lower in the control group than in the experiment group (P < 0.05). At month 1, 3, and 6 of follow-ups, 24 h UPQ of the experiment group was significantly lower than the baseline level (P < 0.01). It was also significantly lower than the level of the control group at the same time point (P < 0.05). There was no significant difference in 24 h UPQ at month 1, 3, and 6 of follow-ups between the control group and the baseline level (P > 0.05). ALB of the experiment group showed an increasing trend. It was significantly higher than the baseline level at month 6 (P < 0.05), which was also higher than that of the control group at same period (P < 0.05). There was no significant difference in the ALB level in the control group (P > 0.05). SCr of two groups showed an increasing trend. SCr of the experiment group was significantly higher at month 1, 3, and 6 follow-ups than the baseline level (P < 0.05). But the increment of SCr was higher in the control group than in the experimental group, and obviously higher than the baseline levels (P < 0.05). eGFR of both groups showed a decreasing trend. The decrement was higher in the control group than in the experimental group (P < 0.05). The proportion of progression of renal functions at month 1, 3, and 6 of follow-ups in the experimental group was 0.0% (0 case), 9.55% (8 cases), and 21.4% (18 cases), while they were 8.3% (7 cases), 21.4% (18 cases), and 40.5% (34 cases) in the control group. There was no statistical difference in the proportion of progression of renal functions between the two groups at month 3 and 6 of follow-ups (P < 0.05). There was no statistical difference in the incidence of adverse reactions between two groups (P > 0.05).

CONCLUSIONQJC could effectively reduce urinary protein of patients with stage 3b DKD, and delay the progression of renal functions.

Adult ; Albumins ; analysis ; Albuminuria ; drug therapy ; Blood Pressure ; drug effects ; Creatinine ; blood ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerular Filtration Rate ; Humans ; Male ; Middle Aged ; Tetrazoles ; therapeutic use ; Treatment Outcome ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan

Objective To retrospectively review the history and development of radiotherapy quality assurance ( QA) in the Affiliated Cancer Hospital of Fudan University, and to report the primary experience and evolvement of an entire QA workflow management. Methods The multidisciplinary QA team has implemented an entire QA workflow management process in the Radiotherapy Center using the failure modes and effects analysis ( FMEA) and plan?do?check?act ( PDCA) tool since April 2015. Treatment data of approximately 6000 patients before and after implementation were compared. Results The error rate was reduced from 17% to 09% after using the entire QA workflow management. Conclusions Entire QA workflow management effectively improves the accuracy and safety of radiotherapy.

Objective:To investigate the differences in efficacy, survival outcomes, and acute and late toxicities for patients with local/regional advanced nasopharyngeal carcinoma (NPC) treated by intensity-modulated radiotherapy (IMRT) in combination with che-motherapy (CT) and by IMRT alone. Methods:A total of 72 newly diagnosed local/regional advanced NPC patients were randomly subjected to IMRT/RT+adjuvant CT (after radiotherapy, RT) (n=42) or IMRT+adjuvant CT (after RT) (n=30). The Kaplan-Meier meth-od was used to analyze the two-year local/regional control rates, distant metastasis-free survivals, and overall survivals. The acute and late radiation toxicities were evaluated based on the toxicity criteria of the Radiation Therapy Oncology Group and European Organiza-tion for Research and Treatment of Cancer. Results:A median follow up period of 13.5 months was included in the study. The one-year and two-year local/regional control rates, distant metastasis-free survivals, and overall survival in the IMRT group were 95.0%, 80.0%, and 95.0%, and 80%, 60.0%, and 75.0%, respectively. For the IMRT+CT group, such rates were 100%, 96.4%, and 96.4%, and 100%, 92.9%, and 92.9%, respectively. The two-year local/regional control rate and distant metastasis-free survivals in the IMRT+CT group were higher than those in the IMRT group (P<0.05). Most patients had grade 1 to grade 2 acute radiation toxicities and grade 0 to grade 1 late radiation toxicities (P>0.05). No patient showed a grade 4 acute or late toxicity. The blood and gastrointestinal toxicity rates were high in the IMRT+CT group (P<0.05). Conclusion:The IMRT+CT treatment has potential advantages over the IMRT in the treatment of local/regional advanced NPC patients in terms of local/regional control and overall survival. The blood and gastrointestinal toxicity rates in the IMRT+CT group were higher than in the IMRT group but still within a tolerable range.

Objective To investigate the efficacy of dexmedetomidine combined with target-controlled infusion (TCI) of propofol and remifentanil for fiberoptic bronchoscopy in the elderly patients.Methods Forty ASA Ⅱ or Ⅲ patients,aged 65-75 yr,with body mass index of 20-30 kg/m2,scheduled for elective fiberoptic bronchoscopy,were randomly divided into 2 groups (n =20 each):control group (group C) and dexmedetomidine group (group D).In group D,a loading dose of dexmedetomidine 0.5/μg/kg was injected at 10 min before induction of anesthesia,followed by infusion at 0.5 μg· kg-1 · h-1 until the end of fiberoptic bronchoscopy.While the equal volume of normal saline was given instead in group C.Anesthesia was induced with TCI of propofol and remifentanil.The target effect-site concentration (Ce) of propofol was 3 μg/ml.When the plasma concentration and Ce were balanced,TCI of remifentanil (target Ce 4 ng/ml) was started.The fiberoptic bronchoscope was placed after consciousness was lost and then the Ces of propofol and remifentanil were adjusted to 1-3 μg/ml and 2-4 ng/ml,respectively.MAP,HR and OAA/S score were recorded before induction (T0),immediately after induction (T1),when the tip of fiberoptic bronchoscope reached the glottis (T2) and carina (T3),at the end of bronchoscopy (T4)and 10 min after the end of bronchoscopy (T5).The consumption of propofol and remifentanil,duration of bron-choscopy,emergence time,adverse cardiovascular events and side effects such as hyoxemia,nausea and vomiting,regurgitation and aspiration were recorded.Results Compared with group C,OAA/S score at T5 and the consumption of propofol and remifentanil was reduced,and emergence time was shortened,and the incidence of hypotension and hyoxemia was decreased in group D (P ＜ 0.05).No patients developed side effects such as hyoxemia,nausea and vomiting,regurgitation and aspiration in both groups.Conclusion Dexmedetomidine (infusion at 0.5 μg·kg-1 ·h-1 after a loading dose of 0.5 μg/kg) combined with TCI of propofol and remifentanil can be safely and effectively used for fiberoptic bronchoscopy in the elderly patients.

ObjectiveTo evaluate the application of chest-abdomen longitudinal diameter ratio,total lung area and lung longitudinal diameter in different gestational ages by two-dimensional ultrasonography.MethodsFive hundred and ninty-two fetus of normal singleton pregnancy between 18 and 40 weeks were ultrasonic scanned by standard 4-chamber view and fetus torso coronal section.The heart area,thoracic area,bilateral lungs pointed to the corresponding diaphragmatic dome gallery on the top of the vertical distance,diaphragmatic dome iterated top to the bottom of the fetal bladder distance were measured respectively.Related regression analysis was done on fitting total lung area and longitudinal diameter with pregnant increasing.ResultsA normal pregnancy fetal total lung area and lung longitudinal diameter were increased as long as the gestational age.The best-fit regression equations of total lung area was:Y =0.83 X +13.894,R2 =0.914;Lung longitudinal diameter was:Y =0.669 X + 3.124,R2 =0.892.Chest-abdomen longitudinal diameter ratio from 18 to 40 weeks gestation was in constant range:0.44 - 0.59.Conclusions Chest-abdomen longitudinal diameter ratio from 18 to 40 weeks gestation is in constant range.The normal pregnancy fetal total lung area and lung longitudinal diameter were increased as long as the gestational age.These parameters may be useful for the prenatal assessment of lung development.