Objective:To study the short-term effects,toxic side effects and immune function of Aidi injection adjuvant GP methods chemotherapy for advanced non-small cell lung cancer(NSCLC).Methods:130 patients with advanced NSCLC were divided into study group and the control group according to treatment methods.The study group were given Aidi injection +GP,and the control group only treated with GP.The short-term effects,toxic side effects gradeⅢ-Ⅳand immune function changes were compared between the two groups.Results:The overall response rate was 40.28%in study group,higher than 37.93% in control group(P>0.05).The two groups showed differences in the rates of leucocyte decrease,gastrointestinal reaction and platelets decrease(P<0.05).The levels of CD4,CD8 and CD4/CD8 didn′t changed obviously in study group(P>0.05),while in the control group the levels of CD4,CD8 and CD4/CD8 changed more significantly than before(P<0.05).The two groups also produced differences in CD4,CD8 and CD4/CD8 after treatment( P<0.05 ) .Conclusion: The short-term effects of Aidi injection adjuvant GP chemotherapy regimen for advanced NSCLC is similar to only GP chemotherapy,but it can obviously reduce the incidence of toxic side effects and prevent the immune func-tion.

Objective To investigate the effects of artesunate (AS) on septic lung injury in mice and to study the modulation of heme oxygenase-1 (HO-1) in lung in order to clarify the mechanism of AS action.Methods Sixty male Kunming mice were randomly (random number) divided into four groups: Sham group (n =15), CLP group (n =15), AS + CLP group (n =15) and AS + ZnPP + CLP group (n =15).Cecal ligation and puncture (CLP) method was employed to induce septic lung injury.AS (15 mg/ kg) was injected into the abdomen of mice 2 hours before the CLP procedures, and ZnPP Ⅸ, an inhibitor of HO-1, was intraperitoneally injected in dose of 40 μmol/kg 1 hour after the AS injection.The equivalent volume of normal saline was intraperitoneally injected instead in mice of Sham group and CLP group.The mice were sacrificed 24 hours after the CLP procedures.The TNF-α, IL-6 in serum were assayed by ELISA method.The lung injury score and wet/dry ratio were measured.The western blotting and immunohistochemistry methods were used to determine HO-1 protein expression in lung tissue.The protein level of nuclear factor-E2-related factor-2 (Nrf-2), an important transcriptional factor of HO-1 in lung tissue was also analyzed by western blotting.One-way analysis of variance (ANOVA) was used for comparisons among the groups, and SNK-q (Student-Newman-Keuls) test was performed for further comparison, and difference was statistically significant at P ＜ 0.05.Results The TNF-α (pg/mL) (54.37 ± 15.59 vs.627.45 ± 117.03, P ＜ 0.05), IL-6 (pg/mL) (81.53 ± 26.89 vs.898.52 ± 222.78, P ＜ 0.05) in serum was increased, and the lung protein exudation, pulmonary edema (wet/dry weight ratio: 4.27 ± 0.22 vs.6.78 ±0.73, P ＜ 0.05), pulmonary pathology injury (lung injury score: 2.20 ± 0.2 vs.13.25 ± 2.67, P ＜ 0.05) were aggravated by CLP.The HO-1 and Nrf-2 were up-regulated in lung tissue in CLP group compared with the sham group (P ＜ 0.05).After the intervention of AS, the HO-1 and Nrf-2 were further increased (P＜0.05), theTNF-α (pg/mL) (627.45 ±117.03 vs.307.88 ±72.33, P＜0.05), IL-6 (pg/mL) (898.52 ± 222.78 vs.413.47 ± 115.14, P ＜ 0.05) in serum, lung protein exudation, pulmonary edema (wet/dry weight ratio: 6.78 ± 0.73 vs.5.05 ± 0.61, P ＜ 0.05), pulmonary pathology injury (lung injury score: 13.25 ± 2.67 vs.4.95 ± 1.46, P ＜ 0.05) were attenuated compared with the CLP group.However, the protective role of AS in the septic lung injury in mice was partly reversed by ZnPP, and no significant difference was detected between the AS + CLP + ZnPP and CLP group (lung injury score: 12.15 ± 2.95 vs.13.25 ± 2.67, P ＞ 0.05;wet/dry weight ratio: 6.78 ± 0.73 vs.6.29 ± 0.82, P ＞ 0.05).Conclusions AS plays protective roles in septic lung injury, and it is attributed to limiting lung inflammation via up-regulation of HO-1.

Objective:To compare the proliferation and osteogenic differentiation between human bone marrow mesenchymal stem cells from orofacial bone(OMMSCs)and those from long bone(BMMSCs).Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.BMMSCs were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method.The surface markers of the cells were detected by flowcytometry.Single-colony formation,CCK assay and cell circle analyses were conducted.Osteogenic differentiation ability was evaluated by ALP activity test and Alizarin red staining after osteogenic induction culture.Results:The cell surface markers STRO-1 and CD105 of both stem cells were positive,CD34,CD31 and CD45 were negative. OMMSCs generated significantly higher numbers of colonies than BMMSCs.In addition,OMMSCs had a higher proliferation rate and more cells in proliferative(S +G2 )stage than BMMSCs.After osteogenic induction for 3,5,7 and 10 d,OMMSCs showed higher levels of ALP activity.OMMSCs formed significantly more mineralized nodules than BMMSCs after 21-day ostogenic induction.Conclusion:The proliferation and osteogenic differentiation capacity of OMMSCs are higher than those of BMMSCs.

OBJECTIVE: To explore the clinical effects of using custom-made tumor prosthesis replacement to treat malignant tumor located in the distal femur or proximal tibia. METHODS: A retrospective study of 29 cases malignant tumors around knee treated surgically from June 2001 to October 2008. In the study there were 19 males and 10 females, aging from 17 to 65 years, with an average of 38.5 years. The location of tumors was at distal femur in 22 cases and 7 cases at proximal tibia. After the tumor was extensively resected or radically resected, custom-made tumor prosthesis replacement was performed for reconstruction, simultaneously, neoadjuvant chemotherapy and radioactive-therapy were used in treating the tumor according the pathology. RESULTS: All patients were followed-up for 18-84 months, with average follow-up period of 54.2 months. The 3 year survival rate was 79.8%, and the 5 year survival rate was 63%. As for the complications, prosthesis breakage occurred in 2 patients, aseptic loosening in 1 patients, 2 had a recurrence of the soft tissue tumor for which the diseased limb amputation was performed. The mean MSTS score showed excellent limb function in 15 patients, good in 8 patients, fair in 5 patients and poor in 1 patient. The overall excellent and good function was obtained in 79% of the patients. No superficial or deep infection occurred. Two patients suffered common peroneal nerve injury, but were cured by symptomatic treatments. CONCLUSION: Application of tumor prosthesis can give a satisfactory functional outcome, which is an effect method in limb salvage treatment of malignant tumor around the knee.

The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rabbits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good biocompatibility and ectopic induced osteogenic activity.
Biocompatible Materials ; Bone Marrow Cells/*cytology ; Bone Matrix/*cytology ; Cells, Cultured ; Chondrocytes/cytology ; Coculture Techniques ; Decalcification Technique ; *Osteogenesis ; Stem Cells/cytology ; Stromal Cells/cytology ; *Tissue Engineering

Objective:To investigate the trait of exosomes serected by mesenchymal stem cells derived form orofacial bone(OMMSCs-Exo) and the communication between the exosomes and macrophages.Methods:OMMSCs were isolated from orthognathic surgical sites and cultured by limited dilution.Their cell surface markers were characterized by flow cytometry.the rate of colony formation and the differentiation potential of OMMSCs were evaluated.Exosomes were prepaired from the culture supernatants of OMMSCs(P4-P6).Transmission electron microscopy(TEM) and western blot were used to identify the exosomes.The expression of miRNAs associated with immunity such as miR-223 and miR-let-7c were determined by Real-time RT-PCR.Human peripheral blood mononuclear cells(PBMCs) were isolated from health donor and co-cultured with OMMSCs-Exo.After co-cultured for 24 h,the communication between exosomes and macrophages was tested using a confocal microscope.Results:Human OMMSCs were proved to have the characteristics of mesenchymal stem cells.The diameter of OMMSCs-Exo ranged from 40 to 160 nm.The OMMSCs-Exo expressed CD63 and CD81 and contained miRNAs associated with immune regulation such as miR223 and miR-let-7c.OMMSCs-Exo could be uptaken by macrophages.After co-culture of OMMSCs-Exo with marcrophages for 72 h,miR223 expression in macrophages increased.Conclusion:OMMSCs-Exo has the potential of immune regulation.

Objective To explore the association of sedentary behavior and time with risk of metabolic syndrome (MS).Methods A total of 10 149 subjects were recruited from local residents aged ≥40 years old in Jiashan County by cluster-random sampling method.The data including physical activity,job,sedentary time,and sleep,etc.were collected.Height and body weight,waist circumference,blood pressure,blood glucose,blood lipid,etc.were determined.Logistic regression was used for correlation analysis.Results The prevalence of MS was 28.64％ in local residents over 40 years old in Jiashan.The rate of sedentary behavior in all subjects was 67.57％,with 3 h sedentary time on average.Compared with non-MS group,the rates of sedentary behavior and sedentary time were significantly higher in MS group (P＜0.01).Multivariate logistic regression analysis showed that sedentary behavior was independently associated with an increased risk of MS after adjustment for age,sex,body mass index (BMI),smoking,drinking,and sedentary time(OR=1.16,95％ CI 1.03-1.30,P=0.017).Increased sedentary time was associated with higher risks of hypertension,type 2 diabetes mellitus,and dyslipidemia (P ＜ 0.05 or P ＜ 0.01).Sedentary time ≥ 5 h/d independently increased the risk of MS (OR =1.15,95％ CI 1.01-1.31,P =0.034).Conclusions There is a high prevalence of MS in adults over 40 years old living in the eastern coastal rural area.MS and its related diseases are closely associated with sedentary behavior and its duration.

A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.

Dysentery, Bacillary ; epidemiology ; Humans ; Molecular Epidemiology ; Shigella ; genetics

OBJECTIVETo study the molecular epidemiology of Vibrio parahaemolyticus isolates from clinical and environmental samples collected in Hangzhou area during 2000 and 2002.

METHODSV. parahaemolyticus isolates from food-poisoning, sporadic diarrhea patients and seafood in market in Hangzhou during 2000 and 2002 were serotyped. From clinical isolates of serotype O3:K6 and environmental isolates of serotype O3:KUT, virulence genes, tdh and trh, were examined by PCR. Molecular typings [including ribotyping, random amplified polymorphic DNA analysis (RAPD), enterobacterial repetitive intergenic consensus PCR (ERIC-PCR)], were also performed.

RESULTSAmong 13 food-poisoning outbreaks caused by V. parahaemolyticus, O3:K6 strains with tdh positive and trh negative were detected in 11 episodes (84.6%), and O4:K8 strains with tdh positive and trh negative were responsible for other 2 outbreaks (15.4%). In 34 V. parahaemolyticus isolates from sporadic diarrhea patients, O3:K6, O4:K8, and O1:KUT strains with tdh positive and trh negative were detected with proportions of 26.5% (9/34), 17.6% (6/34), and 38.2% (13/34), respectively, and other 6 strains were not able to be serotyped. Of 64 isolates from seafood in which both tdh and trh were negative except trh was positive in one O1:KUT strain, 37 belonged to 7 serotypes except O3:K6 or O4:K8, and 9 were not able to be serotyped. The fingprintings of ribotyping, ERIC-PCR, and RAPD showed that almost all of O3:K6 isolates from food-poisoning and sporadic diarrhea patients were genetically close to each other and there were obviously genetical difference between O3:K6 isolates from clinic and O3:KUT isolates from environment.

CONCLUSIONThere was a distinct serotype distribution between V. parahaemolyticus isolates from clinic patients and seafood. One group of close related V. parahaemolyticus O3:K6 strains with tdh positive and trh negative seemed to be prevailing in Hangzhou in those years.

China ; epidemiology ; DNA, Bacterial ; analysis ; Disease Outbreaks ; Food Contamination ; Foodborne Diseases ; Humans ; Molecular Epidemiology ; Polymerase Chain Reaction ; Seafood ; Vibrio parahaemolyticus ; genetics ; isolation & purification