Objective To establish a method of SYBR Green Ⅰ FQ-PCR for detecting the expression of miR-202 in peripheral blood mononuclear cells ( PBMC ) and analyze the expression of miR-202 and its clinical significance in MM.Methods Reverse transcription was performed with specific stemloop primer for miR-202,and then FQ-PCR was used to detected the expression of miR-202 in 21 MM patients and 20 healthy people.Data was presented as mean ± standard deviation ( (x) ± s ).Non-parametric Mann-Whitney test was used to analyze the difference between MM group and control group.In addition,1∶ 125 dilution of one test sample was detected by repeated 5 times,and the same sample was tested one time a day for 3 days,repeated 5 each time.The assessment of repeatability of measurements was done by calculating standard deviation and variation coefficient from threshold cycle (Ct).Results FQ-PCR detection of miR-202 in PBMC was amplified by the standard S-curve,with a single melting curve peak and no complex peak,which showed good specificity.The assay showed good reproducibility (intra-assay coefficient 1.2％ and inter-assay coefficient 3.2％ ) and high sensitivity ( 12.8 pmol/μ1).The expression of miR-202 in MM was 0.014 ( 0.007 - 0.221 ) and 1.844 ( 0.162 - 3.966 ) in normal controls.The expression level of miR-202 was significantly higher in MM than in normal controls (U =48.000,P ＜0.01 ).Conclusions FQ-PCR provide us a rapid,sensitive and specific method for detection of miR-202.The expression level of miR-202 is higher in MM than in normal controls.It is possible to play a role in MM progress and may be a useful marker to evaluate the development and treatment of MM.

Objective To investigate the action and mechanism of NF-κB pathway in up-regulating B cell-activating factor receptor (BAFF-R) expression in multiple myeloma cells induced by IFN-γ.Methods Activated NF-κB were detected with Western blot, while the expression of BAFF-R were measured with RT-PCR and ELISA, and investigated the effect of BAY11-7082 on transcription of BAFF-R mRNA and translation of protein in multiple myeloma cells stimulated by IFN-γ. Results IFN-γ can induce the degradation of IκB-α in time-dependent and dosage-dependent manner, and up-regulated BAFF-R expression in multiple myeloma cells. BAY11-7082, an NF-κB inhibitor, inhibited not only the transcription of BAFF-R mRNA but also the protein of regulated by IFN-γin dosage-dependent manner. Conclusion NFκB may play an important role in high expression of BAFF-R in multiple myeloma cells induced by IFN-γ.

Objective    To introduce the surgical and perioperative strategy for giant mediastinal mass. Methods    The clinical data of 21 patients with giant mediastinal mass who underwent surgical treatment in Xinhua Hospital of Shanghai from January 2007 to July 2016 were retrospectively reviewed. There were 14 males and 7 females, with a mean age of 34.62 ± 22.95 years (range: 11 months to 79 years), and mean weight of 58.07±22.24 kg (range: 10.8 to 90.5 kg). Their clinical manifestation, anesthesia methods, surgical treatment and the prognosis were analyzed. Results    The tumor volume ranged from 8 cm×6 cm×6 cm to 25 cm×25 cm×8 cm. For surgical approach, 12 patients received median sternotomy, 5 anterior lateral incision, 1 posterior lateral incision, 2 "L"-shape sternotomy, 1 cervical and thoracic "]"-shape incision. All patients were given mass radical resection, except one patient with two-stage resection. Twelve patients needed other tissues resection besides the single tomor resection. The operation time was 55-480 (207.86±87.67) min, blood loss volume 700 (10-4 000) ml, intraoperative blood transfusion 800 (0-4 100) ml, postoperative mechanical ventilation time 4.75 (0-87) h, postoperative drainage time 3-12 (7.43±2.66) d, the total drainage volume 295-4 940 (1 584.76±1 173.98) ml, average daily drainage volume 62-494 (204.90±105.76) ml, and postoperative hospital stay 7-47 (11.86±8.51) d. The postoperative complications included pericardial effusion in 1 patient, Horner   syndrome in 1, left recurrent laryngeal nerve injury with the left phrenic nerve injury in 1, right phrenic nerve injury in 1 and delayed wound healing in 1. The remaining patients recovered well. All patients were followed up for 1 month to 9 years. Till September 1, 2016, 5 patients died and 2 suffered recurrent tumor. Conclusion    It is safe to perform surgical treatment after comprehensive evaluation of patients with giant mediastinal mass, perioperative mortality is low, and prognosis in patients with benign tumor is good.

ObjectiveTo investigate the activation of MAPK signal pathway in multiple myeloma and the regulation of BLyS expression levels through MAPK signal pathway; preliminarily study the role of MAPK signal pathway in the up-regulation of BLyS expression levels induced by IFN-γ.MethodsActivated MAPK pathway were detected by Western blot,while the expression of BLyS were detected with RT-PCR and Western blot,and Western blot investigated the effect of MAPK pathway on BLyS expression levels induced by IFN-γ.ResultsIn addition to the expression of ERK,JNK,p38,p-JNK was also expressed in MM cell lines,the MAPK pathway inhibitor targeting JNK SP600125 can down-regulate the expression of BLyS,and its activator anisomycin can up-regulate the expression of BLyS.SP600125 restrained the proliferation and survival of MM cells.ConclusionJNK/SAPK pathway was activated in MM cells; The activated degree of JNK/SAPK pathway and the expression level of BLyS was positively correlated.JNK/SAPK pathway play an important role in the up-regulation of BLyS expression levels induced by IFN-γ.

OBJECTIVETo evaluate the effect of Qufeng Zhidong Recipe (QZR) on the head tic behavior, and the mRNA expressions of Notch1 and dopamine D2 receptor (D2R) in tic disordered mice.

METHODSMouse model like wet-dog shake head tic disorder was established by peritoneal injection of 5-HT2A/C agonist DOI for 14 successive days. The model mice were divided into three groups, the model group, the Chinese medicine (CM) treated group and the Western medicine (WM) treated group, they were intervened respectively with distilled water, QZR (10 g/kg) and haloperidol (1 mg/kg). Besides, a normal control group was set up and gastrogavaged with distilled water. The effect of intervention was evaluated 2 weeks later by estimating the head tic and the creeping distance of animals, and the mRNA expressions of D2R and Notch1 in corpus striatum and prefrontal cortex regions were detected using Real-time PCR.

RESULTSThe wet-dog shake response and the creeping distance of mice were significantly reduced after intervention in both intervened groups, showing insignificant difference between the effects of CM and WM (P > 0.05). The expression of D2R mRNA in corpus striatum was higher than that in the prefrontal cortex (P < 0.01), at the prefrontal cortex, it was 151 +/- 30 in the CM group and 180 +/- 41 in the WM group, and at the corpus striatum, 710 +/- 64 and 850 +/- 80 respectively, all higher than those in the model group (P < 0.05). While the Notch1 mRNA expression in model mice were lower at the prefrontal cortex than at the corpus striatum (P < 0.05). After intervention it was 55 +/- 20 in the CM group and 48 +/- 23 in the WM group at the prefrontal cortex, all significantly lower than that in the model group (P < 0.05).

CONCLUSIONDOI-induced wet-dog shake response could well simulate the clinical characteristics of tic disorder; QZR could improve the tic behavior and creeping distance in the model mice. The up-regulation of D2R mRNA expression after QZR intervention may be related with the down-regulation of Notch1 expression, this findings is worthy of further studies.

Animals ; Corpus Striatum ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Receptor, Notch1 ; genetics ; metabolism ; Receptors, Dopamine D2 ; genetics ; metabolism ; Signal Transduction ; drug effects ; Tic Disorders ; drug therapy

To summarize the clinical experience of treating adults with pectus excavatum by newly modified orthopedic steel plate and Nuss procedure. Methods    The clinical data of 190 adults with pectus excavatum treated by newly modified Nuss procedure in Xinhua Hospital affiliated to Shanghai Jiaotong University School of Medicine from January 2011 to June 2016 were collected. There were 151 males and 39 females aged 21.80±3.96 years ranging from 18 to 45 years. The therapeutic efficacy, Haller index and the lung function index were also analyzed. Results    All patients recovered well after the operation. Two orthopedic steel plates were implanted in 12 patients. Pneumothorax was found in 7 patients postoperatively and 2 of them was treated by puncture extraction. There were 5 patients with poor healing of incision and all of them were healed after the debridement. Other complications such as steel plate shift was found in 2 patients. Both of them recovered after the reoperation. The duration of operation was 36–65 min. The intraoperative blood loss was 5–20 ml. Postoperative hospital stay was 4–7 days. Haller and lung function index improved after the operation (P<0.001). Conclusion    It is effective and safe to treat the pectus excavatum by newly modified orthopedic steel plate and Nuss procedure in adult patients.

Objective    To summarize the experience of right ventricular outflow tract reconstruction with self-made single-valve conduit for the treatment of complex congenital heart disease, and to explore the key points of operation, and to evaluate the short-term and medium-term results of the treatment. Methods    We retrospectively analyzed the clinical data of 65 patients with complex congenital heart disease treated by self-made single-valve conduit from January 2006 to June 2018. There were 42 males and 23 females aged 5-23 (9.9±4.2) years with weight 15-65 (26.2±9.9) kg. There were 19 patients with single valve artificial blood vessel (an artificial vascular group), and 46 patients with single valve bovine pericardium tube (a bovine pericardial tube group). There were 48 patients of ventricular septal defect (VSD) with pulmonary atresia (PA), 10 patients of corrective transposition of great artery with pulmonary artery stenosis, 5 patients of tetralogy of Fallot with single coronary artery malformation, 2 patients of double outlet of right ventricle with pulmonary artery stenosis and single coronary artery malformation. Results    Two patients died early after operation, both of them were VSD/PA patients who underwent radical treatment of extravascular prosthesis. The right ventricular outflow tract pressure difference was 13-37 (25.2±4.9) mm Hg in the artificial vascular group and 5-23 (10.5±3.3) mm Hg in the bovine pericardial tube group. No obvious reflux was found at discharge. The patients were followed up for 8 months to 13 years. One patient was lost. During the follow-up period, there were 5 patients of moderate tricuspid regurgitation, 32 patients of moderate intraductal regurgitation, 7 patients of severe right ventricular outflow  tract obstruction, 11 patients of moderate obstruction and 25 patients of mild obstruction. There was no late death. Heart function classification of all patients was in New York Heart Association classⅠtoⅡ. Conclusion    The self-made single-valve conduit used to reconstruct right ventricular outflow tract shows good clinical effect. Using bovine pericardium tube has less pressure difference of outflow tract in comparison with artificial blood vessel. Bovine pericardium tube is more recommended for young and low weight patients.

Objective    To investigate the perioperative change and the predictive value of myoglobin, creatine kinase-MB (CK-MB), and cardiac troponin I (cTnI) in non-coronary cardiac surgery. Methods    The clinical data of 77 patients undergoing cardiac surgery for non-coronary lesions in the Shanghai Xinhua Hospital from March 2016 to November 2016 were retrospectively reviewed, including 37 males and 40 females with a median age of 2 years. There were simple congenital heart diseases in 45 patients, complicated congenital heart diseases in 10, and heart valve diseases in 22. The levels of myoglobin, CK-MB and cTnI were collected at the first postoperative day. The ventilation duration and the length of ICU stay were recorded. The recovery condition was accessed by senior surgeons. Results    The myoglobin, CK-MB and cTnI concentrations increased at the first postoperative day, and cTnI increased most significantly. The multivariate linear regression analysis indicated that these changes were only related to cardiopulmonary bypass time and aortic cross-clamping time (P<0.001). The high cTnI level was associated with prolonged ventilation duration and length of ICU stay. Fourteen patients (18.2%) did not recovered well, and their cTnI level was significantly higher than that of well-recovered patients (16.8±16.7 ng/ml vs. 5.1±4.4 ng/ml, P<0.001). The cTnI cutoff value of 5.33 ng/ml could predict whether patients had good postoperative recovery (area under the receiver operating characteristic  curve=0.862, P<0.001), and the predictive value of cTnI was superior to that of myoglobin and CK-MB. Conclusion    The increase levels of myoglobin, CK-MB and cTnI post non-coronary cardiac surgery are associated with prolonged cardiopulmonary bypass time and aortic cross-clamping time. cTnI on postoperative 24 h may predict good recovery, and it is a useful biomarker.

Objective    To explore the surgical design and efficacy of novel modified Nuss procedure for pectus excavatum (PE). Methods    We retrospectively analyzed the clinical data of 903 patients with PE who were treated by the new kind of steel bar and the novel modified Nuss procedure. There were 716 males and 187 females at mean age of 2-45 (12.1±6.8) years. Preoperative chest CT scans Haller index (HI) was 3.1-15.2 (4.6±1.3), with 38 patients of mild PE (HI<3.2), 302 patients of moderate PE (HI 3.2-3.5), 521 patients of severe PE (HI 3.6-6.0), and 42 patients of extremely severe PE (HI>6.0). The operative time, operative blood loss, hospital stay time as well as postoperative complications were reviewed and analyzed. Results    All of the 903 patients successfully completed the surgery. The mean operative time was 20-45 (25.2±2.6) min for primary PE and that for special type of PE (including recurring PE, PE patients after heart disease operation and those corrected by 2 bars) was 48-150 (63.5±28.1) min. Blood loss was less than 10 mL for primary PE and 15-50 (23.5±5.5) mL for special type of PE. Postoperative hospital stay was 3-15 (4.5±1.6) d. A total of 845 patients (93.6%) required 1 steel bar insertion, 58 patients (6.4%) required 2 steel bars. Postoperative evaluation of the surgery outcomes revealed the following: excellent in 805 patients, good in 84 patients, fair in 14 patients and poor in 0. The good quality rate was 98.4%. Conclusion    Novel modified Nuss procedure can simplify and optimize the surgical design with good short and mid-term effects.

Objective     To investigate the surgery experience of modified intra/extracardiac conduit total cavopulmonary connection (TCPC). Methods     We retrospectively analyzed clinical data of 47 patients of complex congenital heart disease undergoing intra/extracardiac conduit total cavopulmonary connection in our hospital between January 2008 and December 2015. There were 29 males and 18 females with a median age of 7 years (range 4 to 9 years) and median body weight of 22 kg (range 14 to 38 kg). The heart echocardiography and cardiac imaging confirmed diagnosis suitable for TCPC surgery. Results     There was no early death in the whole group. The mean pulmonary arterial pressure was 16 (12–20) mm Hg and the ventilation time was 14 (7–97) h. The main complications were intractable pleural effusion in 7 patients, low cardiac output syndrome in 3 patients, repeated supraventricular tachycardia in 1 patient. All the patients recovered after treatment. At the end of discharge, the percutaneous oxygen saturation was 85%–96% (92.6%±3.3%). The echocardiography showed the conduit pressure was 0–2 mm Hg. Patients were followed up for 1 to 7 years. Three patients were lost. One patient had intestinal nutrition loss, receving repeated pleural effusion, the treatment was ineffective, died after 4 years. Four patients of repeated pleural effusion improved after treatment. One patient repeated attacks supraventricular tachycardia within 1 year, controlled by amiodaronum, already stopped about 28 months. No recurrence occurred. All survivors were in New York Heart Association (NYHA) functional class Ⅰ or Ⅱ, with good activity tolerance. Conclusion     The modified intra/extracardiac conduit TCPC combines the advantages of both the lateral tunnel and the extracardiac conduit. The operation is simple, used in the treatment of complex congenital heart disease. The short-term and mid-term results are encouraging.