ObjectiveTo investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis in PC12 cells undergone ischemia and hypoxia, and the mechanism involved.Methods PC12 cells at logarithmic phase were collected, and were divided into recombination lentivirus infection group (infected by lentivirus containing HSP70 and fluorescent gene), lentivirus control group (infected by lentivirus containing fluorescin without HSP70 gene) and non-infection group. HSP70 gene and protein expressions in PC12 cells were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot. After being challenged with ischemia and hypoxia for 4 hours, the viability of cells was detected by methyl thiazolyl tetrazolium (MTT), the levels of lactic acid dehydrogenase (LDH) in cell supernatant were determined by LDH measurement test kit. The concentration of intracelluer calcium ([Ca2+]i) was assayed by flow cytometer. The activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and total ATPase were measured by ATPase test kits.Results The expressions of exogenous HSP70 gene and protein were found by RT-PCR and Western Blot in the recombination lentivirus infection group. After being challenged with ischemia and hypoxia, the viability of cells in the recombination lentivirus infection group was increased significantly as compared with the lentivirus control group and non-infection group (A value: 0.575±0.020 vs. 0.395±0.014, 0.363±0.045,t1= 17.996,t2= 10.600, bothP< 0.001), the levels of LDH in culture medium and the concentration of [Ca2+]i were decreased significantly [LDH (U/L): 743.46±23.68 vs. 935.43±34.77, 962.89±26.68,t1= 11.179, t2= 15.044, bothP< 0.001; [Ca2+]i relative fluorescence: (31.60±2.43)% vs. (41.48±3.33)%, (40.40±3.05)%, t1= 5.853,t2= 5.502, bothP< 0.001], and the activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase and total ATPase were increased significantly [Na+-K+-ATPase (μmol·mg-1·h-1): 8.608±0.307 vs. 6.728±0.173, 6.450±0.091,t1=9.237,P1= 0.001,t2= 11.675,P2< 0.001; Ca2+-Mg2+-ATPase (μmol·mg-1·h-1): 10.523±0.036 vs. 7.910±0.209, 8.064±0.195,t1= 9.718,P1= 0.001,t2= 11.535,P2<0.001; total ATPase (μmol·mg-1·h-1): 17.041±0.324 vs. 14.150±0.182, 13.983±0.085,t1= 16.113,t2= 17.602, bothP<0.001]. There was no statistical difference in above indexes between lentivirus control group and non-infection group.Conclusion HSP70 can maintain the PC12 cells calcium homeostasis, which may be one of the important mechanisms of anti-apoptosis.

Objective To evaluate common carotid arterial intima-media thickness (IMT) and elasticity using ultrasound radio-frequency data (RF-data) technology, the normal values of the related parameters were determined in healthy adults for their potential clinical applications.Methods One hundred and sixty healthy adults were included and divided into 4 age groups with 20 males and 20 females in each group.The quantitative parameters of common carotid arterial IMT, common carotid arterial distensibility (CD),local pulse wave velocity(PWVβ) and stiffness(β) were calculated automatically.The correlations that the IMT, PWVβ and β were correlated positively with aging and CD was correlated negatively with between IMT and PWVβ or β, while no correlation between IMT and CD.The positive correlation was found correlation between IMT and age ( P＜0.05) ;In 30～39 and 40～49 years old groups,PWVβ increased and CD decreased with aging;In 50～59 years old group,PWVβ and β increased with aging;But in 60～69 years these parameters changed with aging in subjects＜60 years old.Conclusions RF-data technology could quantitativly evaluate common carotid arterial IMT and elasticity, the normal values of the related parameters were determined with aging,it may be a promising modality to assess the arterial condition.

BACKGROUND:Dragon’s blood is the main ingredient of traditional medicine prescription for promoting granulation, which has been used in clinical treatment of a variety of refractory wounds and achieved the exact effects. But the Dragon’s blood effect on colagen secretion from normal fibroblasts has not been reported. OBJECTIVE: To investigate the effects of Dragon’s blood extract on the proliferation and secret function of fibroblastsin vitro. METHODS: Dragon’s blood was extracted by extracts chloroform, acetoacetic ester, and alcohol in turn. Normal human fibroblasts were respectively cultured in Dragon’s blood extracts of chloroform, acetoacetic ester, and alcohol, DMEM containing 1% dimethyl sulfoxide, and normal culture medium. Then, the fibroblasts were cultured in vitro in different media containing gradient dilutions of Dragon’s blood extracts (0.002, 0.02, 0.2, 2, 20 g/L), which was folowed by cel proliferation determination assessed with MTT assay. Under the optimal concentration, the cel growth curves were drawn and the flow cytometry was used to determine the changes of cel cycle. The concentration of procolagen type III in the supernatant of the fibroblast culture systems was measured by radioimmunoassay. RESULTS AND CONCLUSION:0.2 g/L-2 g/L dilution of Dragon’s blood extracted by acetoacetic ester enhanced the proliferation of fibroblasts in a dose-dependent manner. The 2 g/L was the optimal dilution of Dragon’s blood extracted by acetoacetic ester, and it increased the ratio of S cels in cel cycle than control group and decreased procolagen type III. These findings indicate that Dragon’s blood acetoacetic ester extract can improve the proliferation of cultured human fibroblastsin vitro, and decrease the secretion of procolagen type III of fibroblasts, and it can be beneficial to improve wound healing and inhibit hypertrophic scar.

Objective To investigate antimicrobial resistance,distribution,and carriage of carbapenemase genes of Acinetobacterbaumannii(AB)from two hospitals in Qingdao.Methods 145 AB isolates collected from two hospi-tals (78 from hospital A,67 from hospital B)were performed antimicrobial susceptibility testing,carbapenemase genes were amplified by polymerase chain reaction (PCR);homology analysis were conducted with enterobacterial repetitive intergenic consensus (ERIC)-PCR.Results AB from hospital A were generally resistant to 16 commonly used antimicrobial agents,with the lowest resistant rate of 3.85% to cefoperazone/sulbactam,followed by resist-ance rate of 16.67% to minocycline,resistant rates to the other antimicrobial agents were all>73% . AB from hos-pital B were generally resistant to 23 commonly used antimicrobial agents,but the resistance rates to minocycline and tigecycline were both 0,resistance rates to amikacin and levofloxacin were 23.88% and 38.81% respectively, resistant rates to the other antimicrobial agents were all >64% . All strains carried OXA-5 1 gene,the carriage rates of OXA-23 gene in carbapenem-resistant group were 86.76% (59/68)and 56.67% (34/60)in hospital A and B re-spectively,the difference was significant(χ2= 14.53,P<0.001);OXA-58 gene was detected in 3 isolates in hospi-tal A but not detected from hospital B. 145 AB strains were classified into 8 types,the major prevalence types were type A (n= 71)and E(n= 37);the major prevalence types in hospital A were type A (46.15% )and E(41.03% ), hospital B were type A (52.24% )and C (17.91% ).Conclusion Antimicrobial resistance of clinically isolated AB is serious and prevailed in two hospitals. OXA-23 and OXA-51 genes play an important role in AB resistance to car-bapenems.

OBJECTIVETo investigate the effects of Dragon' s blood extract on proliferation and secret extracellular matrix function of fibroblasts in vitro.

METHODSDragon' s blood was extracted by chloroform, acetoacetic ester, alcohol. Human fibroblast were cultured in vitro in media containing gradient dilutions of Dragon' s blood extracts (0.002, 0.02, 0.2, 2, 20 mg/ml) , which was followed by cell proliferation assessed with MTT assay on 0, 12, 24, 36, 48, 60, 72 h. Under the optimal concentration, the cell growth curves were drawn and the flow cytometry (FCM) was used to determine the changes of cell cycle. On 0, 12, 24, 36, 48, 60, 72 h, the concentration of hyaluronic acid in the supernatant of fibroblast culture was measured by radioimmunoassay.

RESULTS0.2-2 mg/ml Dragon' s blood extracts enhanced the proliferation of fibroblasts in a dose-dependent manner. 2 mg/ml was the optimal dilution of Dragon's blood extract, and it increased the ratio of S cells in cell cycle [(25.80 ± 3.10)%] than control group [(7.50 ± 0.70)%, P < 0.01]. From 12 h to 72 h, in 2 mg/ml Dragon's blood group, concentration of Hyaluronic acid secreted by fibroblasts gradually increased, but were less than control (P < 0.01).

CONCLUSIONSDragon's blood acetoacetic ester extract improved the proliferation of cultured human fibroblasts in vitro, might be beneficial to promote wound healing.

Cell Cycle ; Cell Proliferation ; drug effects ; Culture Media ; chemistry ; Dose-Response Relationship, Drug ; Extracellular Matrix ; Fibroblasts ; cytology ; drug effects ; secretion ; Flow Cytometry ; Humans ; Hyaluronic Acid ; analysis ; secretion ; Plant Extracts ; pharmacology ; Resins, Plant ; Time Factors

Objective To study the biofilm-forming ability and the distribution of biofilm-related genes in Acinetobacter baumannii clinical isolates as well as antimicrobial resistance,to analyze their relationships with the bacterial resistance phenotype.Methods A prospective study was conducted.Biofilm models of 70 strains Acinetobacter baumannii collected in Chengwu County People's Hospital from October 2012 to October 2013 were constructed using 96-well polystyrene plate.In order to analyze the biofilm-forming ability,a qualitative and quantitative analysis was conduct by crystal violet staining assay.And the antimicrobial resistance of different biofilm-forming ability strains was compared including imipenem,amikacin,meropenem,cefepime,sulbactam cefoperazone,trimethoprim,levofloxacin,gentamicin,ciprofloxacin,cefotaxime,ceftizoxime,aztreonam,piperacillin,ceftriaxone,cefuroxime.In addition,the expressions of biofilm-related gene Bap,bfs and intI1 were tested with polymerase chain reaction (PCR) assay.Results Among 70 strains Acinetobacter baumannii,40 strains were multi-drug resistant (57.14％) and 6 strains were pan-drug resistant (8.57％); 68 strains had biofilm-forming ability (97.14％),14 of which were weakly positive,20 were positive and 34 were strongly positive.The antimicrobial resistant rate of Acinetobacter baumannii to imipenem,amikacin,meropenem and cefepime was decreased,it was 30.00％,32.86％,38.57％ and 41.43％,respectively.However,the antimicrobial resistant rates to other commonly used antibiotics were all higher than 50％.The drug resistance of Acinetobacter baumannii to levofloxacin (85.71％,45.00％,38.24％,x2=9.225,P=0.010),cefepime (71.43％,45.00％,29.41％,x2=7.222,P=0.027),gentamicin (78.57％,55.00％,38.24％,x2 =6.601,P=0.037) was significantly decreased when biofilm-forming ability reinforced (weakly positive,positive,hadro-positive).Bap gene positive rate of weakly positive,positive and strong positive biofilm-forming strains Acinetobacter baumannii was 50.00％,65.00％ and 79.41％ (x2=4.244,P=0.120),respectively.Bfs gene positive rate was 35.71％,65.00％ and 88.24％,respectively (x2=13.602,P=0.001) and intI1 gene positive rate was 42.86％,75.00％ and 91.18％,respectively (x2 =12.902,P=0.002).Moreover,the antimicrobial resistances of biofilm-related gene positive strains were higher than the negative,of which the drug resistance of intI1 positive group to amikacin was significantly higher than the negative group (40.38％ vs.11.11％,x 2=5.194,P=0.023).Conclusions The Acinetobacter baumannii collected from the hospital had strong multi-drug resistance as well as strong biofilm-forming ability.The drug resistance of Acinetobacter baumannii decreased when biofilm-forming ability reinforced.In addition,genes,such as Bap,bfs,and intI1,contributed to biofilm formation.

Objective To explore the effect of infrared ray treatment on improving fat granules survival rate and reducing postoperative complications in autologous fat transplantation. Methods The patients who received autologous fat transplantation were divided into infrared ray group and control group.Each group was further divided into breast group and face group. These cases in infrared ray group were radiated for 30 minutes with infrared ray 24 hours later after surgery, once a day for sequential 5 days.No treatment was given to the control group. The survival status and complication were observed and statistically analyzed. Results Infrared ray treatment could increase fat granules survival rate after autologous fat transplantation, but no significant effect was acheived on reducing complications. Conclusion Infrared ray treatment is an effective way to enhance the survival rate of fat granules.

Objective To assess the diagnostic value of ultrasonographic elastography for identifying suspected malignant thyroid nodules. Methods Fifty-one patients with small single solid thyroid nodule underwent two-dimensional and color power Doppler ultrasonography. The size, shape, boundary, internal echo and halo, calcification of thyroid nodule were observed with two-dimensional ultrasound. Then color Doppler ultrasonography was used to detect blood signal and distribution in the nodules. Longitudinal scanning of elastography was performed, and the findings on elastography were classified into Ⅰ-Ⅴ grades. The ultrasonographic results were compared with pathologic findings. Results Six patients were found with grade Ⅰ and Ⅱ benign lesions;9 with grade Ⅲ ( 6 benign and 3 malignant), 36 with Ⅳ and Ⅴ (2 benign and 34 malignant) lesions on ultrasonographic elastography. Taking the elasticity grade Ⅳ or Ⅴ as malignant standard, the diagnostic sensitivity was 91.89%, specificity was 85.71%, accuracy was 90.20%, while the positive and negative predictive value was 94.44% and 80.00%, respectively. Conclusion Real-time ultrasound elastography is a promising imaging technique that can assist in the differential diagnosis of malignant thyroid small single solid nodule.

OBJECTIVETo evaluate the applicability of immune-related response criteria (irRC) in treating non-small cell lung cancer (NSCLC) by Chinese medicine (CM).

METHODSTotally 97 stage III a-IV NSCLC patients were predominantly treated with comprehensive CM. Curative effects were evaluated by three methods such as Response Evaluation Criteria in Solid Tumors (RECIST), Oncologic Curative Effect Evaluation Criteria of Chinese Medicine in Solid Tumor (draft, abbreviated as CM criteria), and irRC. The correspondency and consistency between irRC, RECIST and CM criteria were analyzed and compared. The objectivity of irRC in evaluating curative effect of Chinese medical treatment for NSCLC was assessed.

RESULTSThe correspondency rate of irRC to RECIST was 59. 79% with Kappa value of 0. 379 (U test, P <0. 01). The two criteria had certain correspondence, but with an unsatisfactory consistency. The correspondency rate of irRC to CM criteria rate was 83. 51% with Kappa value of 0.751 (U test, P <0. 01). The two criteria had good correspondence and consistency.

CONCLUSIONSCM criteria had good consistency with CM criteria in evaluating curative effect for Chinese medical treatment of advanced NSCLC. Its results could objectively reflect features and advantages of CM for treating advanced NSCLC.

Asian Continental Ancestry Group ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; immunology ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lung Neoplasms ; drug therapy ; immunology ; Medicine, Chinese Traditional ; standards ; Treatment Outcome

Objective To investigate the clinical significance of anticardiolipin antibodies (ACA)in patients with hemorrhagic fever with renal syndrome(HFRS).Methods The serum ACA- IgM and ACA-IgG in 175 cases of renal diseases and 50 healthy cases were detected by enzyme-linked immunosorbent assay(ELISA). The positive rates of ACA were calculated and compared among different groups.Results The positive rates of ACA-IgM and ACA-IgG in control group were both 0 (0/50),while they were 85.2%(46/54)and 24.1%(13/54)respectively in patients with HERS. 25.0%(13/52)and 21.2%(11/52)respectively in patients with lupus nephritis and 13.0%(9/69) and 8.7%(6/69)respectively in patients with nephrotic syndrome.The poshive rate of ACA lgM in patients with HFRS was higher than that in control group,lupus nephritis group and nephritic syn- drome group(P