AIM: To explore the expression of CD14 in rat Kupffer cells (KCs). METHODS: In rat KCs induced by LPS or the mediators from KCs induced by LPS,the changes of CD14 expression were measured by RT-PCR and immunohistochemistry.The expressions of TNF? mRNA?IL-6 mRNA or the concentrations of TNF??IL-6 were estimated by in situ hybridization and radioimmunoassay,respectively. RESULTS: LPS increased the expression of CD14 in KCs in a dose-dependent fashion (LPS,1 ?g/L-10 mg/L) and in a time-dependent fashion(0.5 h-24 h,peaked at 3-6 hours). While the expression of CD14 in KCs stimulated by the active mediators from KCs which had been exposed to LPS 1 hour were obviously increased. CONCLUSIONS: There was a close relationship between LPS or the active mediators from KCs induced by LPS and the expressions of CD14. It is implied that the increase in CD14 expression may be induced by LPS and the cytokines produced by KCs,it also reveals that there is a auto-regulated loop in CD14 expression.

Objective To discuss diagnosis,treatment and late effects of three patients with acute radiation-induced skin injury after an accident.Methods Medical history collection and physical examination were made in a manner of one or several doctors to one patient.The general health condition was evaluated through laboratory studies,including complete blood count,liver and kidney function,thyroid function and humoral immunity measurement.The genetic analysis of radiation damage was performed by using chromosomal aberrations and micronucleus assay (CB method).The psychological status of the patients was evaluated during medical follow-up.Results Among the three patients,two were diagnosed with degree Ⅲ acute radiation-induced skin injury and one with degree Ⅱ.The medical follow-up showed that two patients had syndrome of neurasthenia.The two cases with degree Ⅲ acute radiation-induced skin injury then became chronic radiation-induced skin damage.Conclusions Acute radiation-induced skin injury could be persistent and become chronic radiation skin damage.It is important for long-term medical follow-up to be taken for patients with acute radiation-induced skin injury.

BACKGROUND:Nano-hydroxyapatite/polyamide artificial bone has good biocompatibility and strong tissue binding force, which exerts a significant role in the recovery of neurological function. OBJECTIVE: To investigate the clinical effect of nano-hydroxyapatite/polyamide artificial bone in anterior cervical fusion. METHODS: Ninety patients with cervical spondylosis and traumatic dislocation of the cervical spine were randomized into observation group and control group. Patients in the two groups underwent nano-hydroxyapatite/polyamide artificial bone and autologous iliac bone implantation, respectively. Surgical condition, Japanese Orthopaedic Association score, vertebral height loss and bone fusion rate were compared and analyzed at different time after implantation. RESULTS AND CONCLUSION:There were no significant differences in surgical time, out of bed time, postoperative hospitalization time between the two groups (P > 0.05), but the intraoperative blood loss was significantly lower in the observation group than the control group (P < 0.05). Japanese Orthopaedic Association scores were significantly increased in the two groups at 3 and 6 months after operation and at last folow-up than before (P < 0.05), but there was no difference in the Japanese Orthopaedic Association score between the two groups (P> 0.05). The vertebral height loss and bone fusion rate had no difference between the two groups at 6 months after operation and at the last folow-up (P > 0.05). No adverse effects occurred in the two groups. These findings indicate that nano-hydroxyapatite/polyamide artificial bone has good clinical effects in anterior cervical fusion, and the treatment effect is equivalent to autologous iliac bone graft.

Objectlve To assess the clinical effects and safety of intravitreal injection of conbercept and macular photocoagulation for non-ischemic macular edema secondary to branch retinal vein occlusion (BRVO).Methods A total of 50 patients (50 eyes) with non-ischemic macular edema following BRVO were retrospectively analyzed.Patients were divided into 2 groups:ICI group (26 eyes) received intravitreal injection of conbercept,laser group (24 eyes) received standard-of-care grid laser for macular edema.The best-corrected visual acuity (BCVA) and central macular thickness (CMT) were observed before and 1 week,1 month,2 months and 3 months after treatment.Then,the changes in pre-treatment and post-treatment were compared,and the related complications were recorded.Results The difference of BCVA before treatment was not significant between two groups (P ＞ 0.05).BCVA at 1 week,1 month,2 months and 3 months after treatment in two groups were all improved,the differences were statistically significant compared with before treatment (all P ＜ 0.05),and the differences were statistically significant between two groups after treatment (all P ＜0.05).At 3 months after treatment,BCVA of 18 patients (69.23％) in ICI group and 8 patients (33.33％) in ICI group improved 2 lines.The difference of CMT before treatment was not significant between two groups (P ＞ 0.05),CMT at 1 week,1 month,2 months and 3 months after treatment in two groups were all decreased,the differences were statistically significant compared with before treatment (all P ＜ 0.05),and the differences were statistically significant between two groups after treatment (all P ＜0.05).13 eyes received repeat intravitreal injection in ICI group,including 2 eyes at 1 month,7 eyes at 2 months with CMT ＞250 μm,and 3 eyes at 3 months.No severe side effect related with drug and intravitreal injection occurred in the two groups.Conclusion Mean BCVA change and CMT change are significantly greater in the intravitreal injection of conbercept than the standard-of-care grid laser group for the macuiar edema secondary to non-ischemic BRVO.

Objective To investigate the immunological effects of thymoglobulin (RATG) on human CD4+and CD8+cells for costimulatory molecule gene expression and the production ofimmune-regulatory cytokines. Methods CD4+and CI8+T cells were isolated and purified fromnormal human peripheral blood mononuclear cells (PBMC) followed by incubation with RATG at37℃. Cells and culture supematants were collected at 24, 48, and 72 h after incubation, and analyzedby real-time quantitative polymerase chain reaction (RT-PCR) for CTLA-4, CD154, forkhead box P3(Foxp3), OX40, IFN-γ, IL-2, IL-10 and CD25 gene expression, and multiplex cytokine detectionassay for IFN-y, IL-2, IL-10, and IL-4 production. Untreated and rabbit isotype Ig-treated cells wereused as negative controls. Results RT-PCR demonstrated that RATG pre-treated CI+and CD8+cells upregulated the expression of CTLA-4, OX40, Foxp3, CD25, IFN-γ, IL-10 and IL-2 genes, anda dramatic increase of supernatant IFN-γ, IL-10, IL-2 and IL-4 was revealed 24 h after treatment asdetermined by multiplex cytokine detection assay when compared with negative controls. Theupre gulation of CTLA-4, Foxp3, OX40, IL-10 and CD25 was reduced, and a down-regulation ofCD154 and IL-2 gene expression was revealed 48 h after treatment. Cells, treated with RATG for 72h, demonstrated up-regulation of CTLA-4, Foxp3, OX40, IFN-y and CD25 gene expression, and theexpression of IL-2 and IL-10 genes was down-regulated. Additionally, supernatant IFN-γ, IL-2,IL-10 and IL-4 levels were decreased. Conclusion RATG stimulates CI4/CD8 T cells to up-regulatecostimulatory molecules and release immune regulation associated cytokines IF'N-γ, IL-2, IL-10in vitro. These results suggest that the unique effect of RATG on CD4+CD8+T cells may be animportant mechanism for its action in inducing immunoregulation, immunosuppression and transplanttolerance.

Objective To explore the expression and the role of monocyte-derived costimulatory molecuels during xenogeneic immune responses. Methods Porcine endothelial cells (PEC) were isolated from aorta, and subcultures were performed. CD4+ cells and monocytes were purified from human peripheral blood mononuclear cells (PBMC). PBMC-PEC co-cultures were established, and the cells were collected followed by staining with florescent-labeled monoclonal antibodies and analyzing by FACS. In selected experiments, monoclonal antibodies specific for CD154, CD80 and CD86 were added into PBMC-PEC co-cultures, and the effects of co-stimulatory molecule blockade in inhibiting lymphocyte proliferation in response to PEC were determined by 3H-thymidine up-take. The proliferation of CD4+ cells induced by PEC-conditioned monocytes with or without co-stimulation blockade was evaluated. Results PBMC-PEC co-incubation demonstrated dramatic lymphocyte proliferation as determined by 3H-thymidine up-take. FACS found that resting monocytes expressed only CD86 but not CD40 and CD80. CD14+ monocytes from PEC-stimulated PBMC demonstrated up-regulation of CD80 and CD40 expression. The up-regulation of CD86 was revealed. PEC-activated monocytes induced CD4+ cell proliferation while resting monocytes did not and this proliferation was inhibited by anti-CD154, anti-CD80 or anti-CD86 antibodies. Conclusions CD14+ monocytes play an important role during xenogeneie immune responses in indirect antigen presentation and co-stimulation- The interaction between monocyte-derived co-stimulatory molecules and CD4+ cell-derived CD154 and CD28 delivers secondary signal and induces CD4+ proliferation, and the co-stimulation blockade inhibits xe-nogeneic cell-mediated immune responses.

Objective Delayed xenograft rejection (DXR) is a major barrier to the long-term xenograft survial.This study evaluated the interaction between human peripheral blood mononuelear cells (PBMC) and porcine endothelial cells (PEC),and the effects of new generation of rabbit antihuman leukocyte polyclonal antibody (newRALG) inhibiting xenogeneic cell-mediated immune responses.Methods newRALG was obtained from rabbits after immunization with activated lymphocytes and monoeytes.PEC were isolated from aorta,and human PBMC were isolated from peripheral blood.Co-cultures of PKH-26 labeled PEC with PBMC were established,newRALG,thymoglobulin,isotype Ig and scavenger receptor (SR) ligand poly G were added into the co-cultures.Cells were collected,then FACS analysis was carried out to detect the up-take of PEC membrane by monocytes and the expression of costimulatory molecules.Lymphocyte proliferative responses to PEC with or without antibody were evaluated by a xenogeneie mixed lymphocyte-endothelial cell reaction (xMLER).Results FACS analysis revealed that monocytes from PBMC-PEC co-cultures became positive for PKH-26 following their interaction with PKH-26 labeled PEC,indicating that they engulfed PEC membranes during activation.PKH-26 positive monocytes up-regulated the CD40 and CD80 expression.Furthermore,SR blockade with poly-G prevented PEC membrane up-take by monocytes,newRALG greatly reduced SR-mediated PEC membrane up-take.The effects of thymoglobulin in inhibiting PEC membrane uptake were limited.xMLER demonstrated strong lymphocyte proliferation in response to PEC,and lymphocyte proliferation was dramatically inhibited by newRALG but not isotype Ig at a dosmdependent manner.Conclusions Monocytes play an important role in xenogeneic immune responses.SR ligand poly G inhibits PEC membrane up-take.newRALG inhibits PEC membrane up-take by monocytes,suggesting that newRALG blocks SR.Additionally,newRALG inhibits lymphocyte proliferation in response to PEC.These results suggest that this new polyclonal preparation may thus impair the initiation of xeno-specific immune responses and prevent xenograft rejection.

BACKGROUND:Differentiation of bone marrow mesenchymal stem cels is induced by integrated factors.In vitro interaction of cytokine complex and certain cel mechanical stimulation is carried out to further improve the efficiency of bone marrow mesenchymal stem cels differentiating into nucleus pulposus-like cels. OBJECTIVE:To investigate the differentiation of bone marrow mesenchymal stem cels into nucleus pulposus-like cels induced by transforming growth factor-β1 and insulin-like growth factor-1 under hydrostatic pressure. METHODS: Bone marrow mesenchymal stem cels from adult rats were separated, cultured and purified in vitro. Passage 3 cels were induced in vitrowith transforming growth factor-β1 and insulin-like growth factor-1 under hydrostatic pressure (hydrostatic pressure group), with transforming growth factor-β1 and insulin-like growth factor-1 under normal pressure (drug group), or with normal culture medium under normal pressure (blank control group). RESULTS AND CONCLUSION:At day 14 after culture, polygonal nucleus pulposus-like cels were observed in the hydrostatic pressure group, but irregular cels in the drug group. There was no obvious change in the blank control group. Levels of colagen type II and DNA were higher in the hydrostatic pressure group than the other two groups. These findings indicate that the combination of transforming growth factor-β1 and insulin-like growth factor-1 can successfuly induce the differentiation of bone marrow mesenchymal stem cels into nucleus pulposus-like cels under hydrostatic pressure, and the differentiation efficiency is higher under hydrostatic pressure than under normal pressure.

Objective To investigate the cachexia morbidity among hospitalized patients with digestive system cancer and evaluate its impact on clinical outcomes.Method By analyzing the clinical data of 5 118 hospitalized patients with digestive system cancer in Zhongshan Hospital,Fudan University from January 2012 to December 2013,we investigated the cachexia morbidity and compared the clinical outcome between cachectic patients and noncachectic patients.Results The overall cachexia morbidity of hospitalized patients with digestive system cancer was 15.7％ (803/5 118).The highest cachexia morbidity was 34.0％ (89/173),found in patients with pancreatic cancer.In cachectic group and non-cachectic group,the overall completion rate of radical resection was 67.1％ (539/803) and 74.5％ (3 214/4 315),respectively (P =0.000).Compared to the non-cachectic group,the cachetic group had significantly longer postoperative hospital days [(11.5 ±6.2) d vs (9.4 ±4.9) d,P =0.003],slower postoperative recovery of bowel function [(3.4 ±0.9) d vs (3.2 ±0.8) d,P =0.013],longer postoperative time to intake semifluid [(4.4 ± 1.5) d vs (3.9 ± 1.3) d,P =0.002],and more postoperative complications in 28 days after surgery [8.9％ (48/539) vs 5.8％ (186/3 214),P=0.006].After surgery,131 patients in the cachectic group were transferred to the ICU,and 646 patients in non-cachectic group transferred to the ICU (24.3％ vs 20.0％,P=0.026).Compared to the non-cachecic group,the reoperation rate [3.2％ (17/539) vs 1.5％ (48/3214)],ventilator support rate [8.0％ (43/539) vs 5.7％ (184/3 214)],and mortality [2.4％ (13/539) vs 1.1％ (35/3 214)] of the cachectic group were all significantly higher (P =0.006,0.042,0.011).Conclusions Cachexia is common in hospitalized patients with digestive system cancer,especially in patients with pancreatic cancer.Cachexia has negative impact on the clinical outcomes.

Objective: To observe the characteristics of serum homocysteine (Hcy) distribution and the epidemiological characteristics of high serum level of Hcy (HHcy) in Kazakh population.
Methods: The stratiifed cluster random sampling method was used to investigate 1003 kazakh individuals in Xinyuan county of Xinjiang including 433 male and 570 female, and they were individuals into 5 age groups:①(25-34) years, n=333,②(35-44) years, n=306, ③(45-54) years, n=230, ④(55-65) years, n=90, ⑤>65 years, n=44. The questionnaire survey, physical check-up and blood biochemical examination were performed and compared among different groups.
Results: ① The geometrical mean of Hcy was 13.3μmol/L, and the male was higher than female as 16.0μmol/L vs 11.6μmol/L. ②By age stratiifcation, the serum levels of Hcy elevated with the increased age accordingly. Except for those with the age>65 years, the serum levels of Hcy were all higher in male than female in other 4 age groups. ③The average rate of HHcy prevalence was 31.5%, and the male was higher than female as 49.9%vs 17.5%.④The average rate of H type hypertension prevalence was 35.1%, and the male was higher than female as 44.3%vs 28.1%. There were 87.6%of H type hypertensive patients with H type hypertension, and the male was higher than female as 95.0%vs 80.0%, all P<0.001.
Conclusion: The serum levels of Hcy were different from gender and age; the prevalences of HHcy and H type hypertension were higher in Kazakh population at Xinyuan county of Xinjiang. Community intervention should be conducted to improve the public health condition in Kazakh population.