BACKGROUND:To improve the survival rate of transplanted hematopoietic stem cells, dynamic monitoring of plasma content of human cytomegalovirus (HCMV) and rapid screening of early active HCMV infection in hematopoietic stem celltranplantation recipients, thus to guide the clinical medication, is preferred. OBJECTIVE:To evaluate the usefulness of fluorescence quantitative PCR assay for early detection of HCMV activation after hematopoietic stem celltransplantation. METHODS:Real-time fluorescence quantitative PCR assay was applied for HCMV monitoring in 656 blood samples from 41 hematopoietic stem celltranplantation recipients and 60 control blood samples. RESULTS AND CONCLUSION:In 656 blood samples, 96 samples were positive, and the HCMV DNA copies ranged from 5.0×102 to 1.0×107 IU/mL. Timely initiation of therapy resulted in the rapid clearance of DNA-viraemia but it recurrenced in short time by drug-resistent. Two cases from 12 postive recipients died from HCMV infection. The quantitative detection of HCMV DNA by real-time PCR is a rapid method for monitoring HCMV infection and the early diagnosis in patients after hematopoietic stem celltransplantation.

Objective To investigate the feasibility of dose verification of helical tomotherapy (HT) using the PTW Seven29 2D-ARRAY with Octavius phantom.Methods The 12 patients HT plan were verified with the PTW Seven29 2D-ARRAY combining Octavius phantom.The detector array was guided and registered by MVCT imaging,and measured dose distributions on the corresponding planes were compared with those calculated by the treatment planning system (TPS).Based on several different Gamma criteria,the Gamma analysis method was utilized to evaluate the dose verification.Results According to the different acceptance criteria combination of dose difference/distance to agreement at different threshold level including 3％/2 mm,3％/3 mm,3％/4 mm,3％/5 mm at TH5,TH10,TH20,TH30,the mean passing rates with γparameter ≤ 1 were all above 91.7％ and 93.9％ when PTW Seven29 2D-ARRAY was horizontal and vertical.The dose distribution measured by the 2D-ARRAY combining Octavius phantom was well consistent with that calculated by the TPS.Conclusions 2D-ARRAY with Octavius phantom can get satisfying results when used in HT plan dose verification and is a viable tool to replace traditional film dosimetry system in clinical quality assurance work.

Objective To evaluate the feasibility, safety and effectiveness of interventional occlusion with thin waist occluder for perimembranous ventricular septal defect with pseudoaneurysm. Methods During the period from March 2008 to March 2013 at Shanghai Yodak Cardiothoracic Hospital , interventional occlusion with thin waist occluder was carried out in a total of 96 patients with perimembranous ventricular septal defect (VSD) complicated by pseudoaneurysm. The patient’s age varied from 2 years to 36 years. Before the procedure, routine left ventricular angiography and ascending aortic angiography were carried out. Left ventricular angiography and ascending aortic angiography as well as the transthoracic ultrasonography were performed immediately after the procedure to observe the therapeutic results. Follow-up checkups with transthoracic ultrasonography, EKG and chest X-ray film were conducted at one week as well as at one, 3, 6 and 9 months after the surgery were conducted. Results Successful occlusion of VSD was obtained in 93 patients, with a success rate of 96.8% (93/96). After the surgery, small amount of shunt flow was detected in 8 patients, which disappeared within 48 hours in 7 patients, and in the other one the shunt flow disappeared one month later which was confirmed by follow-up examination. After the treatment, complete right bundle branch block occurred in 4 patients, complete left bundle branch block in 5 patients, and transientⅢdegree atrioventricular block in one patient. The conduction block disappeared at the time of discharge in all patients. The left ventricular end-diastolic diameter (LVEDD) decreased from preoperative (47.5 ± 5.2) mm to postoperative (45.2 ± 5.1) mm, the difference was statistically significant (t = 18.33, P < 0.01). During the follow-up period lasting for 9 months, no delayed cardiac arrhythmia, valve damage or sudden death occurred. Conclusion For the treatment of perimembranous ventricular septal defect with pseudoaneurysm, interventional occlusion with thin waist occluder is safe and effective with excellent mid-term effect. Based on the imaging findings, including the location, shape, ruptured orifice, number of the perimembranous pseudoaneurysm as well as the distance between the pseudoaneurysm and the aortic valve, to select suitable occluder and proper occlusion site is the key to ensure a successful surgery.

Objective To analyze the response rate and prognostic factors for patients with locally recurrent rectal cancer treated with hypofractionated chemoradiotherapy without reresection.Methods Totally 52 patients with locally recurrent rectal cancer received hypofractionated irradiation and concurrent chemotherapy from January 2006 to January 2013 were enrolled.All patients received intensity-modulated radiotherapy (IMRT).The median dose was 63.4 Gy (61.6-64.4 Gy) at 2.2-2.3 Gy/f,5 f/week.Thirteen patients underwent prophylactic irradiation at lymph nodes region,the total dose of 45-50.4 Gy with conventional fraction and a simultaneous integrated boost was used.All patients received concurrent chemotherapy,capecitabine at 1 650 mg·m-2 ·d-1,divided into 2 times,5 d/week.The variables were compared by the chi-square test or Fisher's exact test.Local control (LC) and overall survival (OS) were calculated with using the Kaplan-Meier method.Results For all patients,the clinical complete response (CR),partial response (PR),stable disease (SD) and progressive disease (PD) was 23.1％,38.5％,32.7％ and 5.8％,respectively.The response rate (CR + PR) for patients with previous irradiation to pelvis and without were 37.1％ and 71.1％,respectively (x2 =5.40,P ＜ 0.05);for patients with 1 and 2 or more recurrent subsites were 81.8％ and 46.7％,respectively (x2 =6.63,P ＜ 0.05).Acute grade 3 skin and hematologic toxicities occurred in 19 patients (36.5％) and 1 patient (1.9％),respectively.None occurred grade 4 toxicity and none occurred grade 3 or more gastrointestinal and urologic toxicities.Four patients showed severe late toxicity of anastomotic stricture and performed a stoma at transverse colon.No other severe late toxicities were observed.The LC at 5 years was 49.1％ and the OS was 23.1％.Conclusions For patients with locally recurrent rectal cancer,hypofractionated chemoradiotherapy without resection is an acceptable and effective regimen,the response rate and long-term outcomes are promising.

Objective To investigate the feasibility of dose verification of intensity modulated planning of helical tomotherapy (HT) using three-dimensional semiconductor array (Delta4) and find a more time-consuming and accurate method to validate the delivery dose.Methods Delta4 detector array was used to verify the HT plan dose distribution of 10 patients.The precise setup of detector array was guided and registered by MVCT imaging.After the implementation of delivery,the measured dose distributions on the corresponding planes were compared with those calculated by the treatment planning system (TPS).According to the different acceptance criteria combination of dose difference or distance to agreement at different threshold level including 3％/2 mm,3％/3 mm,3％/4 mm,3％/5 mm at TH5,TH10,TH20,TH30,TH70,TH90,the γ analysis method was utilized to evaluate the dose verification.Results The dose distribution measured by the Delta4 was well consistent with that calculated by the TPS.The mean γpassing rates were all above 94.89％.Conclusions Delta4 detector array can get satisfying results when used in HT plan dose verification and is a viable tool to replace traditional film dosimetry system in clinical quality assurance work.

OBJECTIVE: To explore the application of double pedicle blood supply pectoralis major myocutaneous flap in reconstruction of laryngeal and hypo pharyngeal defect after carcinoma resection. METHOD: All the patients received initial treatment. Pectoralis major myocutaneous flap was Applied in one-stage reconstruction in 22 cases of laryngocarcinoma and hypopharyngeal carcinoma with post-operation defect. During operation, pectoral branches of thoracoacromial vessels and branch of lateral thoracic vessels were reserved and prepared to be double pedicle vessel fascia bundle without muscle at the beginning part, rearched to defect area through subclavian tunnel. After operation, it is necessary to closely observe the subcutaneous swelling, quantity and color of the drain and others of neck surgery area to exclude the occurrence of surgical hematoma. When there is suspicion, do timely exploration. RESULT: Pectoralis major myocutaneous flaps of 22 patients were all survived. Pharyngeal fistula occurred in 2 cases and was cured by change of dressing. Twenty-one cases received radiotherapy after operation without secondary flap necrosis. After follow-up of 6 months to 24 months, one case of roll repairment of circumferential defect in cervical esophagus,found to have anastomotic stenosis 8 months after radiotherapy and then underwent gastrostomy treatment,the other 21 cases showed good recovery of swallowing function. Among 13 cases with laryngeal function reserved, only 1 case wearing tube and the extubation rate was 92.3% without communication barriers, no patient died in the short follow-up period. CONCLUSION For double pedicle blood supply pectoralis major nyocutaneous flap, blood supply are fully guaranteed and does not require complicated equipment to observe and monitor the flap blood supply after operation, it is important to exclude factors that may lead to muscular flap necrosis to ensure the survival of pectoralis major myocutaneous flap.
Esophagus ; surgery ; Fistula ; pathology ; Humans ; Hypopharyngeal Neoplasms ; surgery ; Laryngeal Neoplasms ; surgery ; Larynx ; surgery ; Myocutaneous Flap ; Pharynx ; pathology ; surgery ; Reconstructive Surgical Procedures

Objective To evaluate the clinical value of multiplex ligation-dependent probe amplification (MLPA) technique used in karyotype analysis of chorionic villi from missed abortion. Methods Feb 2008 to Oct 2008, 91 patients with missed abortion diagnosed by hormonal measurement, type B ultrasound and physical exam matched with 20 normal pregnant women undergoing artificial abortion were enrolled in this study. Chorionic villi was obtained by suction dilation and curettage in aseptic condition, then those villi was cultured and analyzed by traditional cytogenetic karyotyping method, in the mean time, the DNA extracted from villi was detected by MLPA. The results of chromosomal G-banding of chorionic villi were compared between two methods. Results The diagnostic concordance of MLPA and traditional karyotyping was observed in 92% (84/91) cases, there were 84 cases in the case group with diagnostic concordance by traditional karyotyping and MLPA except 7 cases of euploidy could not be detected by MLPA. The 84 cases included 40 normal karyotype,29 trisomy of euchromosome, 1 double trisomy of euchromosome, 10 monosomy X , 1 monosomy X combined with trisomy of euchromosome, 2 chimaera of X chromosome, 1 structural abnormity of euchromosome. Among 7 cases with discordance diagnosis, 2 cases with trisomy and 5 cases with tetrasomy of euchromosome were identified in traditional karyotyping, however, they were all diagnosed with normal disomy by MLPA. Of 20 villi from normal pregnancy, two methods got the consistent results. Conclusion The MLPA was rapid and efficacy method used for analyzing aneuploids in chorionic villi.

AIM:To observe the effect of Wnt/β-catenin signaling pathway on diabetic ulcer.METHODS:Diabetic animal model was established in the female Wistar rats by intraperitoneal injection of low-dose streptozotocin fol-lowing high-fat diet feeding.A circular wound was made on the dorsum of the rats in both control group and diabetic group. The condition of wound healing was recorded and the structures of the wound tissues were observed by HE staining in the 2 groups at 3, 7 and 14 d after wounding.The expression ofβ-catenin, GSK-3βand Rspo-3 at mRNA and protein levels in the wound tissues was detected by RT-PCR and ELISA.RESULTS:In diabetic group, the wound healing rate was lower (P<0.05), and the inflammatory cells, fibroblast cells and new capillaries in the wound tissues were fewer than those in control group.The expression of β-catenin and Rspo-3 at mRNA and protein levels in the wound tissues in control group was significantly higher than those in diabetic group, and the expression of GSK-3βwas exactly the opposite (P<0.05). CONCLUSION:The down-regulation of Wnt/β-catenin probably resultes from the decreased level of Rspo-3, which may be one of the reasons for delaying the diabetic ulcer healing.

Objective To study the intracellular localization and temporal expression of CPSIT_0271 in Chlamydia psittaci-infected cells; and to investigate the effects of recombinant GST-CPSIT_0271 protein on the expression of proinflammatory cytokines including IL-6, IL-1βand TNF-αby THP-1 cells.Methods The gene encoding CPSIT_0271 of Chlamydia psittaci was expressed as fusion protein ( GST-CPSIT_0271 ) in E.coli.The polyclonal antibody was prepared by immunizing BALB /c mice with the purified recombinant pro-tein.Antibody titer was determined by ELISA .Indirect immunofluorescence assay ( IFA) was performed to lo-cate the endogenous CPSIT_0271 protein in C.psittaci-infected cells .The expression characteristics of CPSIT_0271 protein were detected by Western blot in C.psittaci-infected HeLa cells at different time points .The levels of IL-6, IL-1βand TNF-αwere analyzed by ELISA after stimulating THP-1 cells with different concentrations of CPSIT_0271 protein.Results CPSIT_0271 protein was found to express in the chlamydia inclusion of C.psittaci-infected HeLa cells .The expression of CPSIT_0271 protein was detected firstly at 36 h and increased at 48 h after C.psittaci infection.The titer of anti-CPSIT_0271 specific antibody in GST-CPSIT_0271 immu-nized mice reached to 1 ∶16 000.GST-CPSIT_0271 protein increased the levels of IL-6, IL-1βand TNF-αin THP-1 cells in a dose-dependent manner in the range of 2 to 5 μg/ml.The levels of TNF-αand IL-1βreached their peaks at 24 h, and IL-6 level peaked at 48 h upon the stimulation by 5 μg/ml of GST-CPSIT_0271 pro-tein.Conclusion CPSIT_0271 expressed in inclusion bodies of Chlamydia psittaci in the infected cells , sug-gesting it might be a late expression gene .GST-CPSIT_0271 protein shows good immunogenicity and enhances the expressions of IL-6, IL-1βand TNF-αin THP-1 cells.