Diaphragm is the principal respiratory muscle.Diaphragm dysfunction resulting from mechanical ventilation,chronic cardiopulmonary disease,medicine,cardio-thoracic surgery,septic shock and chronic malnutrition predisposes to weaning failure from mechanical ventilation and prolongs intensive care unit (ICU) stay.Traditionally,the method of assessment of diaphragm structure and function was invasive,lacking specific,and was difficult to be used bedside.A number of recent studies have been suggested that ultrasonographic evaluation has been proposed as an accurate,safe and noninvasive method of quantification of diaphragm structure and function.This article reviews the related researches of technique and the clinical applications of ultrasonography in the evaluation of diaphragmatic structure and function,which may possibly result in timely identification of patients with diaphragm dysfunction and improve the assessment of recovery from diaphragm weakness.

Objective To investigate the morphological characteristics and proliferation ability of diabetes mellitus-derived adipose-derived stem cells (ADSCs) by comparing with normal-derived ADSCs.Methods The ADSCs could be achieved from diabetes mellitus' adipose tissue by the similar method of normal ADSCs isolation and culture,and then the differnce was compared between the two groups about their morphologies under microscope,and the proliferation ability of two groups was determined by CCK-8.Results ADSCs in patients with diabetes were obvious hypertrophy and irregular in morphology.A values of ADSCs in diabetes and non-diabetes were 0.210+0.002 and 0.390+0.006 in 1st day;0.250+0.015 and 0.443+0.023 in 2nd day;0.368+0.014 and 0.726+0.033 in 3rd day;0.368+0.014 and 0.726+0.033 in 4th day;1.767+0.072 and 3.153+0.067 in 5th day and 1.810+0.072and 3.170+0.021 in 6th day,respectively.The difference was statstically significant at the beginning from 4th day (P＜0.05).Conclusions There is obvious difference of the microscopic morphology between the diabetes mellitus-derived ADSCs and normal-derived ADSCs.The proliferation ability of the diabetes mellitus-derived ADSCs is lower than normal-derived ADSCs.These difference could be closely related to ulcerous non-healing wounds in patients with diabetes mellitus.

Objective To evaluate the effects of radioimmunoimaging with 99Tcm-EGFR-McAb or 99Tcm-CD44-McAb or combined application of both on nude mice bearing lung adenocarcinoma. Methods Animal model of nude mice bearing lung adenocarcinoma was established. The direct labeling method of 99Tcm was applied to labeling monoclonal antibodies for EGFR and CD44, and then the properties of the labeling antibodies purified by SephadexG50 Column were identified. The radio-immuno-image as well as body distribution concerning nude mice bearing lung adenocarcinoma was studied by application of 99Tcm-EGFR-McAb or 99Tcm-CD44-McAb alone or combined application of both. Results The labeling rates of 99Tcm for EGFR-McAb and CD44-McAb were (91.5?3.8)% and (92.3?4.1)% respectively. Specific activities of the labeled antibodies were (2.8?0.3)MBq/g and (2.9?0.5)MBq/g, respectively. The rates of radiochemical purity were (96.5?2.8)% and (96.2?3.1)%, respectively. The tumor tissue had high intake of the two labeled antibodies according to radioimmunoimaging result. The radioactivity concentration of the combined application of the two labeled antibodies was obviously higher than that of the single application. The T/NT relative values measured through ROI technique were 5.58?0.46, 2.72?0.22, and 2.30?0.18, respectively. The body distribution result of the labeled antibodies and their imaging results were basically identical. Conclusion The optimal target and non-target ratios could be obtained by application of 99Tcm-EGFR-McAb or 99Tcm-CD44-McAb alone in nude mice bearing lung adenocarcinoma. The target and non-target ratios could be enhanced through the combined application of the two antibodies.

Objective To investigate the diagnostic values of serum free fatty acids (FFA) and urine podocalyxin (PCX) in the patients with type 2 diabetic nephropathy(DN).Methods A total of 120 patients with type 2 diabetes mellitus (DM) were enrolled,and they were divided into three groups,normal albumin group(NA),microalbuminuria group(MA) and heavy albuminuria group(HA),based on the urinary albumin-creatinine ratio(UACR).In addition,40 healthy persons participated in medical examination were selected as the control group.Serum FFA levels were detected by a biochemical analyzer and urine PCX levels by ELISA.The correlation between them was analyzed by Pearson correlation,and their diagnostic values in type 2 DN were evaluated by the receiver operating characteristic curve (ROC).Results Serum FFA levels in the NA,MA,HA and control groups were (0.61 ± 0.14),(0.81 ± 0.13),(0.95 ± 0.18) and (0.49 ± 0.11) mmol/L,respectively,and urine PCX levels were (1.86 ± 0.45),(4.47 ± 1.48),(6.72 ± 1.40)and(1.38 ±0.24) ng/mL,respectively.The levels of serum FFA and urine PCX in type 2 DM patients were significantly higher than those in the control group(P ＜ 0.05),and increased with the progression of type 2 DM.Pearson correlation analysis showed that serum FFA levels were positively correlated with urine PCX levels(r =0.73,P ＜ 0.05).The sensitivity of serum FFA,urine PCX and combined detection in the diagnosis of type 2 DN were 74.1％,80.6％ and 89.5％,respectively,and the latter significantly higher than the former two (P ＜ 0.05).Conclusion Serum FFA and urine PCX levels increase significantly in diabetic patients with renal injury,and both of them have important clinical values in the diagnosis of type 2 DN.

Objective To investigate the safety and effect of high intensity focused ultrasound ablation(HIFU) on residual liver cancer in patients after radiotherapy. Methods Twenty liver cancer patients with twenty five residual tumors after three-dimensional conformal radiotherapy or stereotactic radiotherapy received HIFU ablation. Liver function and periphery blood cell counts were performed before HIFU and at 1 week after HIFU in all patients. Enhanced MRI and α-fetoprotein (AFP) level were performed before HIFU and at 2 weeks after HIFU to evaluate the effect of HIFU ablation. The survival of all patients was assessed by the Kaplan-Meier method. Results The mean follow-up time was (12.6 ± 8.0)months. The median survival time and 1-year survival rate were 22 months and 87.5% respectively. No skin burns were observed in all patients. As compared with before HIFU,there were no significant differences in the levels of albumin,alanine transarninase, aspartate transarninase, total bilirubin, direct bilirubin, blood red cell counts,blood white cell counts and blood platelet counts at 1 week after HIFU (paired t test, P =0. 156,0. 356,0. 203,0.659,0. 531,0. 519,0. 310,0. 346, respectively). Significant difference in AFP level of 9 patients with AFP＞20 μg/L was observed before HIFU and 2 weeks after HIFU (paired t test, P =0.030). Among 25 residual liver tumors,sixteen with complete ablation and six with ablation volume of ≥80% were observed by enhanced MRI at 2 weeks after HIFU. Conclusions HIFU may be a safe and effective new method to ablate residual liver cancer after three-dimensional conformal radiotherapy or stereotactic radiotherapy.

Objective Analysis of the expression of CD38,CD133 antigen and their clinical significance in myelodysplastic syndrome (MDS).Methods CD38 and CD133 antigen were analyzed by flow cytometry in 31 cases of MDS patients.Results CD38 was expressed in 18 cases (58.1% ),among them,12 cases were found to be myelodysplastic syndrome refractoryanermia ( MDS-RA ),accounting for 57.1%,6 cases were found to be MDS-RAEB,accounting for 66.7%.CD133 was expressed in 20 cases(64.5% ) ,among them,11 cases were found to be MDS-RA ( 52.4% ),1 case MDS-RAS,and 8 cases of MDS-RAEB,accounting for 88.9% .CD38 expressed significantly higher in MDS than anemia and relatively normal group ( P ＜ 0.05 ).CD133 expression in anemia groups was different from MDS-RA without statistical significance ( P ＞ 0.05 ),but was significantly different from relatively normal group (P ＜0.05).CD133 expression was significantly higher in these with MDS-RAEB than those in anemia and normal group ( P ＜ 0.05 ).Conclusions Combining with conventional antibodies,flow cytometry used in detection of CD38 ,CD133 ,could improve the diagnostic rate of MDS.

Objective To investigate the effect of calpeptin on diaphragmatic injury and atrophy under controlled mechanical ventilation in rats.Methods A total of 24 SPF Sprague-Dawley (SD) rats were randomly divided into anesthetized control group (CON group),24-hour controlled mechanical ventilation group (CMV group),and 24-hour CMV + treatment with calpeptin group (CMVC group),with 8 rats in each group.Animals in the CON group received an intraperitoneal injection of pentobarbital sodium without CMV and continuous infusion of pentobarbital sodium.A small-animal ventilator was used for 24 hours in rats of CMV group.Rats of CMVC were treated with a specific calpain inhibitor calpeptin (4 mg/kg).The drug was injected subcutaneously 2 hours before and 8,15 and 23 hours after mechanical ventilation.Changes in diaphragm ultrastructure,light microscopic picture,and myosin heavy chain (MHC) expression were observed.Results ① Alignment of myofilaments and normal Z-band,and the shape of mitochondria were maintained in CON group as revealed by electron microscope.The signs of misalignment of myofibrils,disruption of Z-band and vacuolar mitochondria were found in CMV group,and they were obviously improved in CMVC group.The density of muscle injury (× 10-2/μm2) in CMV group was significantly higher than that in control group (36.8 ± 13.7 vs.6.4 ± 6.3,t=6.373,P=0.001),and that in CMVC group was significantly lowered (17.6 ± 9.1 vs.36.8 ± 13.7,t=3.694,P=0.002).② In CON group,the diaphragm fibers appeared regular in cross section without pathologic change under light microscopy.Fuzzy muscle striations,irregular muscle fibers,centralized nuclei and swelling of capillary endothelial cells were observed in CMV group,while pathological changes in the CMVC group were milder significantly.③ In CMV group,the density of MHCslow and MHCfast was lower compared with that of CON group,and the gray value was lowered by 61.1％ (t=8.138,P=0.001) and 77.1％ (t=8.844,P=0.001),respectively,especially in MHCfast.However,the gray values of MHCslow and MHCfast were increased by 1.51 folds (t=4.601,P=0.010),and 1.33 folds (t=2.859,P=0.011),respectively,after treatment with calpeptin,and the elevation was more significantly in MHCslow.Conclusions Diaphragmatic injury and atrophy were found after CMV for 24 hour.Calpeptin could reverse the detrimental effects of CMV,and it suggested that calpain plays an important role in modulating the ventilator-induced dysfunction of the diaphragm.

ObjectiveTo investigate the role of lymphocyte apoptosis and endoplasmic reticulum stress (ERS) on the development of sepsis and their association with the prognosis of sepsis patients.Methods A prospective cohort study was conducted. Seventy septic patients admitted to intensive care unit (ICU) of Shanghai East Hospital of Tongji University were enrolled. Blood samples were collected on days 1, 3 and 7 to measure percentage of circulating apoptotic lymphocyte with flow cytometry analysis. The relative expressions of endoplasmic reticulum specific glucose regulated protein 78 (GRP78) mRNA and transcription factor CHOP mRNA were measured by real-time reverse transcription-polymerase chain reaction (RT-PCR). The correlation between CHOP mRNA expression and percentage of circulating apoptotic lymphocyte was analyzed by Spearman relative analysis. The patients were divided into death (n = 23) and survival subgroups (n = 47). Twenty healthy volunteers during the same period were selected as the healthy control group.Results① Rate of lymphocyte apoptosis: compared with healthy control group [(2.86±0.66)%], septic patients, either survival or death subgroup, exhibited higher rate of lymphocyte apoptosis on days 1, 3 and 7 [survival subgroup: (12.44±4.43)%, (8.57±3.38)%, (6.78±3.35)%; death subgroup: (14.42±2.01)%, (11.32±2.53)%, (8.87±3.62)%, allP< 0.01], and it was obvious on day 1, and the phenomenon became less marked gradually. The rate of circulating apoptotic lymphocytes did not differ between the death and survival subgroups on day 1, but there was a significant difference in the rate on day 3 and day 7 (bothP< 0.05).② The expression of CHOP mRNA (2-ΔΔCt):compared with that in healthy controls [(2.56±1.09)×10-3], CHOP mRNA expression was increased on days 1, 3 and 7 in septic patients [survival subgroup: (5.83±1.96)×10-3, (4.24±1.60)×10-3, (4.15±1.64)×10-3, death subgroup:(37.20±20.70)×10-3, (18.80±13.90)×10-3, (9.28±7.78)×10-3, allP< 0.01], and it was more obvious in the death subgroup, as it was increased by 6.38, 4.43, and 2.24 folds (P values was 0.000, 0.000, and 0.001), but it decreased rapidly in death subgroup.③ The expression of GRP78 mRNA (2-ΔΔCt): compared with healthy controls [(3.31±2.04 )×10-3], the expression of GRP78 mRNA in both survival and death subgroups increased in septic patients on day 1 [(5.83±2.00)×10-3, (11.30±6.48)×10-3, bothP< 0.01], and they decreased subsequently. The expression of GRP78 mRNA in the survival subgroup declined to the levels of the healthy control group on day 3 and day 7 [3 days: (3.99±1.60)×10-3, 7 days: (3.30±1.35)×10-3, bothP> 0.05], and GRP78 mRNA expression in the death subgroup was gradually lowered, but it was still higher than that in the healthy control group [3 days: (7.27± 3.64)×10-3, 7 days: (5.23±1.94)×10-3, bothP< 0.01].④ Spearman relative analysis showed that the expression of CHOP mRNA was positively correlated with the rate of lymphocyte apoptosis (r = 0.414,P = 0.000 ).Conclusion The increase in the rate of lymphocyte apoptosis and activation of ERS play an important role in the development of sepsis, and it is associated with worse outcome in the septic patients.

Objective To explore the expression of Metastasis-associated in colon cancer-1 (MACC1) in patients with nasopharyngeal carcinoma (NPC), and its relationship with clinical pathologic characteristics and prognosis Methods The expression of MACC1 was detected in 130 cases of NPC and the relationship among the MACC1 expressions, clinical pathologic characteristics and prognosis of NPC was analyzed. Results Positive expression rate of MACC1was 68.5% in the NPC and MACC1 expression was associated with advanced T stages, lymph node metastasis and advanced clinical stages of NPC (P < 0.05). The results of Kaplan-Meier survival analysis showed that the five year overall survival rate in patients with positive expressions of MACC1 (45.9%) was significantly lower than that of those with negative expressions (73.7%) and the difference was statistically significant (P < 0.05), Cox multi-factor analysis results showed that MACC1 expression was an independent prognostic factor for NPC (P = 0.004). Conclusion MACC1 abnormal expression is closely related to the invasion and metastasis of NPC and it is expected to become a new target for gene therapy of NPC.

Objective To explore the relationship of expression and the effect of preoperative radiotherapy of endothelin-1 (ET-1) and pyruvate kinase M-2 (PKM2) in rectal carcinoma.Methods Immunohistochemical method was used to detect the expression of ET-1 and PKM2 proteins of rectal cancer tissues in 96 cases.The expressions of ET-1 and PKM2 were analyzed with the effect of preoperative radiotherapy in rectal cancer tissue.Results The high expression of ET-1 protein was 59 cases (61.46％).The high expression of PKM2 proteins was 54 cases (56.25％).The high expressions of ET-1 and PKM2 protein were worsen the effect of tumor regressive grade (TRG) than lower expressions of those after preoperative radiotherapy of rectal cancer tissue (P ＜ 0.05).The protein expression of ET-1 and PKM2 were positively correlated (P =0.006).Conclusions The high expressed ET-1 and PKM2 proteins in rectal cancer are closely related to preoperative radiotherapy resistance.ET-1 and PKM2 proteins are expected to become new targets of radiotherapy sensitivity and radiotherapy sensitization of rectal cancer.