ObjectiveTo investigate the role of caspase 3 in HMME-induced apoptosis in hypertrophic scar fibroblasts (HSFs).MethodsFibroblasts were obtained from 10 patients with untreated hypertrophic scar,and subjected to a primary culture.After 4 to 6 passages of culture,the HSFs were divided into 3 groups to remain untreated(control group),be treated with HMME followed by photodynamic therapy (HMME-PDT group),or the combination of HMME and Z-DEVD-FMK followed by photodynamic therapy (caspase 3 inhibitor group).At 12 hours after the therapy,HSFs were collected and immunofluorescence microscopy was used to observe the fluorescence intensity of caspase 3 after staining with fluorescein isocyanate (FITC) and popodium iodide (PI),flow cytometry was performed to determine the percentage of caspase 3-positive HSFs and apoptosis rate in HSFs after single staining with FITC and PI respectively.Results The fluorescence intensity of caspase 3 was weak in the control group and caspase 3 inhibitor group,but was strong in the HMME-PDT group.An increased percentage of caspase 3-positive HSFs was noted in the HMMEPDT group compared with the control group and caspase 3 inhibitor group(30.86％ ± 1.21％ vs.3.12％ ±0.28％ and 2.46％ ± 0.18％,t =19.92,21.76,both P ＜ 0.05).The apoptosis rate in HSFs was significantly higher in the HMME-PDT group and caspase 3 inhibitor group than in the control group(30.54％ ± 3.78％ and 10.46％ ± 2.15％ vs.2.45％ ± 0.22％,t =35.90,27.97,both P＜ 0.05),and higher in the HMME-PDT group than in the caspase 3 inhibitor group.ConclusionsThe apoptosis in HSFs induced by HMME-PDT is closely related to the activation of caspase 3,while caspase 3 seems to be dispensable for the apoptosis.

Objective To investigate the correlation between chronic insomniacs' sleep quality and age,gender,education level,anxiety,depression and sympathetic skin response (SSR) in chronic insomniacs.Methods General information of 197 outpatients with chronic insomnia was recorded,including age,gender and education,etc.They were tested by Pittsburgh's Sleep Quality Index (PSQI),Hamilton's Anxiety Scale (14 item version) (HAMA14),Hamilton's Depression Scale (24 item version)(HAMD24) and Sympathetic Skin Response (SSR).Distribution properties of different age,gender and education groups were studied.Chronic insomniacs were divided into mild insomnia group (7 ≤ PSQI ＜ 14)and moderate-severe insomnia group (PSQI ≥ 14).Dependency relation analysis and stepwise linear regression analysis were conducted among indices of PSQI scores,HAMA14 scores and total score,HAMD24 scores and total score,SSR positive incidence.Results Among 197 chronic insomniacs (male,50 cases,25.4％;female,147 cases,74.6％),insomniacs aged over 40 accounted for 77.2％.Female patients were older than male patients with statistical significance,of whom those aged 40-60 years accounted for the highest proportion of 37.1％.Female patients with less education (junior high school and below)accounted for the highest proportion of 50.3％ (73/197),whose education level was generally lower than male patients.Among 197 chronic insomniacs,104 cases (52.8％,99/197) had mild insomnia and 93cases (47.2％) had moderate-severe insomnia.Total score of HAMA14 of patients with moderate-severe insomnia was significantly higher than that of patients with mild insomnia (16.47 ± 5.40 vs 12.51 ± 4.53;t =5.552,P＜0.01).There was statistically significanct difference in subitem HAMA14 scores of anxiety somatization factor (4.31 ± 2.26 vs 5.90-3.10,t =5.600,P ＜ 0.01) and spiritualized anxiety factor (10.5 ± 72.97 vs 8.20 ± 3.00,t =4.157,P ＜ 0.01) between mild and moderate-severe groups with insomnia.Total score of HAMD24 of patients with moderate-severe insomnia was significantly higher than that of patients with mild insomnia (18.04 ± 5.91 vs 13.41 ± 5.05;t=3.931,P＜ 0.01).There was statistically significanct difference in scores of most HAMD24 subitems including anxiety/somatization (3.56 ± 1.51 vs 2.94 ± 1.28;t =3.110,P =0.002),cognitive dysfunction (2.91 ± 1.68 vs 2.17 ± 1.57;t=3.191,P=0.002),retardation (2.331 ±1.31 vs 1.72 ±1.22;t=3.939,P=0.01),dyssomnia (4.51 ± 1.54 vs 3.01 ± 1.80;t =6.228,P ＜0.01) and hopelessness factor (2.29 ± 1.46 vs 1.66 ± 1.07,t =3.459,P =0.001;except body weight and diurnal variation factor) between groups with different degrees of insomnia.SSR abnormal incidences of moderate-severe insomniacs were significantly higher than that of mild insomniacs.The proportion of poorly differentiated waveform and not elicited waveform in SSR abnormal groups had statistically significant difference.The Pearson correlation analysis showed that PSQI scores in chronic insomnia patients and HAMA14,HAMD24 score as well as abnormal rate of SSR were positively correlated (r =0.439,0.465,0.249,all P ＜ 0.01).Conclusions Chronic insomnia was commonly seen in middle-aged women with education level of junor high school and below.The degree of sleep quality and anxiety,depression as well as the abnormal rate of SSR was positively correlated in patients with chronic insomnia.

Objective To evaluate the cause of ischemia related to myocardial bridge (MB) by using SPECT/CT MPI and CTCA.Methods A total of 294 patients with chest pain,tightness or palpitation undergoing both CTCA and MPI were retrospectively enrolled in this study from March 2008 to March 2013.Among them,49 patients (26 males,23 females,age:32-85 (55.4± 16.6) years) had MB.Locations of MB and myocardial ischemia were recorded.Fused MPI/CTCA was analyzed.If there was no mural atherosclerotic plaque-related stenosis on CAG at the same location of coronary artery where ischemic myocardium was found,then MB was considered as the ischemic cause.Myocardial ischemia rates of different MB locations were compared by x2 test.Results Among 49 patients with MB,3 cases had MB in proximal segment of LAD,34 in mid LAD,4 in distal LAD,3 in septal branch,2 in distal LCX,1 in intermedius,and 2 in mid RCA.There were 41 cases with myocardial ischemia.Myocardial ischemia in 32 cases was caused by MB,including 23 caused by MB in mid LAD.The myocardial ischemia rates of the most common MB location (mid LAD,n =34) and other locations (n =15) were not significantly different (67.6％ (23/34) vs 60.0％ (9/15),x2 =0.27,P＞0.05).Conclusions MB is commonly found in the mid LAD.The myocardial ischemia rates caused by MB is not related the MB location.Hybrid MPI/CTCA could evaluate the sites of coronary MB and myocardial ischemia simultaneously and therefore may be useful to evaluate the relationship between MB and myocardial ischemia.

Objective To investigate the prognostic significance of follicular lymphoma international prognostic index (FLIPI) and FLIPI2 in Chinese FL patients for better prognostic model.Methods 63 newly diagnosed FL patients who received rituximab plus CHOP (cyclophosphamide,doxorubicin,vincristine and prednisone)-like chemotherapy from January 2003 to December 2010 were retrospectively analyzed according to FLIPI and FLIPI2 scoring system.Results According to FLIPI,31 patients (49.2 ％) were stratified into the low risk group (0-1 point),24 (38.1％) into the intermediate risk group (2 points),and 8 (12.7 ％) into the high risk group (3-5 points).As for FLIPI2 scoring,17 cases (27.0 ％) were classified in low risk group (0 point),42 cases (68.3 ％) in intermediate risk group (1-2 points),and 4 cases in high risk group (3-5 points).The overall response rates and 2-year progression free survival (PFS) rates of the three risk groups were 93.5 ％ (29/31),91.7 ％ (22/24),75.0 ％ (6/8) (P =0.27),and 85.7 ％,75.2 ％,58.8 ％ (Log-rank =1.063,P =0.59) respectively in FLIPI.In terms of FLIPI2,the overall response rates and 2-year PFS rates of the three groups were 100.0 ％ (17/17),90.7 ％ (39/42),50.0 ％ (2/4) (P =0.01) and 92.9 ％,74.0 ％,33.3 ％ (Log-rank =7.075,P =0.03).Conclusion FLIPI2 presents more robust prognostic significance than FLIPI in disease progression perspective for FL patients treated with R-CHOP like regimens in this study.

Objective To analyze the main clinical features of human infection with H7N9 avian influenza complicated by acute respiratory distress syndrome (ARDS).Methods A retrospective analysis of complete clinical data of 9 cases of human infection with H7N9 avian influenza complicated by ARDS admitted from March 2013 to December 2014 admitted to Department of Critical Care Medicine of the Affiliated Drum Tower Hospital of Nanjing University Medical School was conducted.Results Nine patients' mean age was (46.3±12.3) years, male accounting for 66.7% (6/9). The main clinical features: ① In the duration of acute phase, high fever, cough, hemoptysis sputum, reduction of white blood cell count (WBC) and platelet count (PLT), and myocardial enzyme elevation were the features of the disease. Chest CT showed pulmonary consolidation and ground-glass like shadows. ② On admission, all their oxygenation indexes (PaO2/FiO2) were less than 200 mmHg (1 mmHg = 0.133 kPa) of which 66.7% (6/9) was less than 100 mmHg, mean (98.9±62.8) mmHg; 55.6% (5/9) required invasive mechanical ventilatory support; 77.8% (7/9) combined with shock, and hemodynamic monitoring showed peripheral vascular resistance was decreased. ③ There were secondary bloodstream infection in 5 cases and lung infection in 4 cases, accounting for 77.8% (7/9). ④ In 22.2% (2/9) patients, the virus relapsed after the anti-virus therapy was stopped for 7 days, then immediately antiviral treatment was used again, it was still effective.Conclusion Human infection with H7N9 avian influenza complicated by ARDS has typical clinical symptoms, laboratory tests and imaging features, often associated with distributive shock, and at the late stage, secondary pulmonary or blood infection and the virus resurgence may occur.

Objective:To investigate the effects of 125I radioactive particle implantation in the treatment of maxillofacial malignancy. Methods:43 patients with maxillofacial malignancy were treated with 125I radioactive particle implantation.The procedure was carried out according to the treatment planning system(TPS),with the particles spaced uniformly at 1 cm intervals and with the activity of 0.7 m Ciby 41 particles per case on a verage.All patients were followed up for 6-60 months.Results:The whole treatment procedurewas successful,and no particle displaced.The follow-up rate was 93.02% and treatment effective rate(CR+PR) was 90.70%.Norecur-rence was foundinall target areas.The mortality due to tumor was 9.30%and total survival rate was 74.43%.The cumulative survival rate of the patients in 0.5,1,2,3 and 5 years was 93.0%,85.7%,79.3,69.8% and 56.9% respectively.Survival periodon aver-age was 36.06-50.04 months,with the median of 43.05 months.The longest tumor-free survival period was 60 months.Radiation in-jury rate was 20.93% and only level 1 radiation injury was observedinall the cases.Facial nerve dys function was found in 2 cases and recovered after treatment.Conclusion:Treatment of maxillofacial malignancy by implantation of 125I particles is convenient and mini-mally invasive.The treatment canincrease survival rate of the patients and guaran tee the oropharynxes' function.

Objective To observe the phosphorylation of Smad3 in hyperplastic scar fibroblasts (HSFs) induced by hematoporphyrin monomerthyl ether (HMME) followed by photodynamic therapy (PDT).Methods Fibroblasts were isolated from the hypertrophic scar tissues of 10 patients and subjected to culture in vitro.After 3-5 passages,the HSFs were divided into 4 groups:control group receiving no treatment,PDT group pretreated with HMME of 4 μg/ml followed by PDT,HMME group induced by HMME alone,and laser group irradiated with laser alone.Fluorescence microscopy was used to observe the expression of Smad3 after immunofluorescent staining with anti-Smad3 antibody,and Western blot to detect the expression of Smad3 and phosphorylated Smad3 in these HSFs.Paired t test was conducted to compare the difference in Smad3 and phosphorylated Smad3 expression between these groups.Results The total fluorescence intensity of Smad3 was similar between these groups,but the intranuclear fluorescence signal was significantly weaker in the PDT group than in the control group.The level of phosphorylated Smad3 was statistically decreased in the PDT group compared with the control group (0.20 ± 0.02 vs.0.92 ± 0.15,P ＜ 0.05),but no significant difference was observed between the HMME group and laser group (P ＞ 0.05).Conclusion PDT may inhibit the proliferation of HSFs via attenuating the phosphorylation of Smad3.

Objective To investigate the role and mechanism of B7-H4, a negative costimulatory molecule, in mediating the immunomodulatory effects of mesenchymal stem cells C3H10T1/2 (C3H10) on T cell polarization. Methods The lentiviral vectors that carried the shRNA targeting mouse B7-H4 were transfected into mouse mesenchymal stem cells (C3H10-B7-H4). The cells were co-cultured with PHA-acti-vated mice spleen lymphocytes before and after the transfection. ELISA was performed to detect the concen-trations of cytokines in supernatants of cell culture in order to elucidate the effects of B7-H4 expressed by C3H10 on T cell polarization. A mouse model of experimental allergic encephalitis (EAE) was established. Fifty C57BL/6 mice were divided into five groups including control group, EAE group, C3H10 group (injec-ting EAE mice with C3H10 cells), C3H10-NC group ( injecting EAE mice with C3H10-NC cells) and C3H10-B7-H4 group (injecting EAE mice with C3H10-B7-H4 cells). ELISA was performed to detect the soluble form of IL-2, IL-17, IFN-γ and IL-4 in plasma samples. Results Knocking down the B7-H4 gene with shRNA significantly decreased the expression of B7-H4 on C3H10 cells, which weakened the inhibitory effects of C3H10 cells on the secretion of IL-2, IL-17 and IFN-γ by spleen lymphocytes. The therapeutic effects of C3H10-B7-H4 cells on mice with EAE were weakened after silencing the B7-H4 gene expression, which was manifested as higher nerve function score and earlier onset and bring forwarded peak time of EAE than those of the C3H10 group. Treating EAE mice with C3H10-B7-H4 cells was less efficient in inhibiting the expression of IL-2, IL-17 and IFN-γin plasma. However, knocking down the B7-H4 gene had no signif-icant effect on the expression of IL-4 in terms of treating EAE with C3H10 cells. Conclusion The co-inhib-itor molecule B7-H4 expressed on C3H10 cells mediated the treatment of EAE with C3H10 cells by regula-ting Th1 and Th17 effector T cells.

Objective To evaluate vascular surgery and interventional technique applied in Takayasu's arteritis. Methods Data of 26 patients of Takayasu's arteritis admitted between January 2006 and December 2009 were retrospectively analyzed. The sex ratio(M/F) was 1: 4. 2, age averaged at (27±15)y. There were 16 cases of type Ⅰ , 7 cases of type Ⅱ and 3 cases of type Ⅲ according to Lupi-Herrera classification. 25 patients received surgery including 16 patients undergoing pecutaneous transluminal angioplasty operations, 9 patients doing traditional bypass surgery, and one patient was treated conservatively. Results 23 case-times of percutaneous transluminal angioplasty (PTA) were performed in 16 patients, including 12 cases of balloon angioplasty and 4 cases of stent angioplasty. Four significantly stenotic and occluded carotid arteries were revascularized successfully in 5 patients. Thrombosis of the carotid artery was found in one patient after balloon angioplasty. There were 4 patients in which repeated PTA treatment up to a total of 11 times were needed to guarantee vessel patency. Open surgery succeeded in 9 patients, and clinical symptoms were relieved in all cases during peri-operative period. 22 patients were followed up for 12 -46 months,one patient died of cerebral hemorrhage 3 months post-operation, one patient was found pseudoaneurysm at anastomotic stoma, and 2 patients suffered from anastomotic restenosis.Conclusions Vascular surgery played important role in the therapy of Takayasu's arteritis. PTA can be used repeatedly. Surgical bypass operation is difficult in technology, and can be used in cases that fail to response to PTA or in patients with severe cerebral ischemia.

OBJECTIVETo compare therapeutic effects between suspine rotation and flexion manipulation for treating lumbar disc herniation.

METHODSFrom December 2008 to January 2011, 134 patients with lumbar disc herniation were treated, including 74 males and 60 females,ranging in age from 17 to 67 years,33 patients with age over 50 years old. According to diagnosis the ladder of the 9 cases bulging type, 74 hernia type, 51 cases free type. The patients were randomly divided into treatment group ( 69 cases) and control group (65 cases). All the patients were treated with the three-dimensional computer-controlled traction therapeutic apparatus,with continued traction for 20 minutes. After traction, flexion manipulation and rotat-ing manipulation were respectively adopted in treatment group and control group (on alternate days one time, 3 times as a course of treatment). The symptoms and signs (including back pain and discomfort, lower limb pain and numbness, powerless urination and defecation,numbness in perineum, straight-leg raising degree,ability of lower extremity walking,work and live) of patients were observed after treatment.

RESULTSAll patients were followed up for 0.5 to 6 months with an average of 1.1 months. After treatment, the symptoms and signs of patients have significantly improved (P < 0.01), but the symptomes of powerless urination and defecation,numbness in perineum was not improved obviously (P>0.05). Forty-two of 69 patients in treatment group got excellent result, moderate in 20, poor in 7; Twenty of 65 patients in control group got excellent result, moderate in 25,poor in 20.

CONCLUSIONThe whole effect of lumbar backwards rotation manipulation is more satisfactory than flexsion manipulation for treating lumbar intervertebral disc herniation. But spinning force and times should be noticed.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Intervertebral Disc Displacement ; physiopathology ; therapy ; Lumbar Vertebrae ; physiopathology ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Recovery of Function ; Rotation ; Young Adult