BACKGROUND: Presently, the biomaterial used in ligament tissue engineering such as collagen protein, polylactic acid, polyglycolic acid, small intestinal submucosa, glycan and nanomaterial are characterized by rapid degradation, resulting in inflammatory reaction after applying in host.OBJECTIVE: To investigate mechanical strength and in vitro degradation of silk scaffold and explore the reaction to macrophages.DESIGN: Controlled experiment.SETTING: Experiments were performed at the Department of Orthopaedics, Union Hospital, Huazhong University of Science and Technology from September 2004 to January 2005.MATERIALS: White raw Bombyx mori silkworm fibers of size 20/22 (according to the manufacturer) were obtained from the market. Bundles of 30 parallel fibers were prepared for a bundle of scaffold, which was put into fervens 5g/L Na2CO3for degumming. Ratio of Na2CO3 solution (Ml) to raw silk (g) was 1000.METHODS: In vitro degradation: 8cm long silk scaffold was weighed after drying. Subsequently, the silk scaffold was separately dipped into phosphate buffer saline (PBS) and 1.0g/L collagenase prepared with PBS. Twelve weeks later, silk scaffold was weighed to calculate weight loss rate. Simultaneously, tensile test was performed to detect the ultimate tensile strength (UTS) of samples. Culture of monocyte strain RAW264.7:2×108L-1 macrophage suspension (1mL) were separately added in a silk scaffold group, a control group and a lipopolysaccharide (LPS) group. At days 1 and 7, cell supernatant was collected from each group. Tumor necrosis factor-α(TNF-α) levels were measured by enzyme linked immunosorbent assay (ELISA).MAIN OUTCOME MEASURES: ① Changes in weight loss rate and UTS of the silk matrices after incubated with collagenase and the PBS. ②TNF-αlevels in the supernatant of each groups at days 1 and 7.RESULTS: Mass of silk matrices reduced by over 50% after incubated with collagenase for 8 weeks, but no change was found in PBS. UTS decreased by over 50% 8 weeks after incubated with collagenase, but no change was detected in PBS. At days 1 and 7, TNF-α levels in the supernatant was less in the silk scaffold group; TNF-α levels in the supernatant was significantly higher in the LPS group than in the silk scaffold group (P<0.01), but no significant difference in TNF-α levels was measured between the silk scaffold group and the control group (P>0.05).CONCLUSION: After 12-weeks degradation, silk scaffold still has good mechanical properties. Macrophages possess immunological inertia at days 1 and 7 after inoculated with macrophages.

Objective To establish a quantitative method of ferulic acid by semi-bionic extraction ( SBE ) coupled with high performance liquid chromatography ,and compare the validity of ferulic acid in Qibaipingfei capsule by water extraction ( WE) method and SBE method .Methods The separation was carried out on a Welch-C18 column (250mm ×4.6mm,5μm).A mixture of methanol-1%(V/V)acetic acid solution(33.67)was used as the mobile phase at a flow rate of 1.0 mL/min in isocratic elution mode .The column temperature was kept at 35 °C, and the detection wavelength was set at 315 nm.Results The calibration curve was linear ( r =0.999 8, n =6 ) in the range of 0.013 4 ~0.134 0mg/mL of ferulic acid.The recoveries were 98.93% with RSDs of 0.53%. Conclusion On the basis of the advantages of simplicity and good reproducibility ,this method can be used for the determination of ferulic acid ,and also we can concluded that abstraction of Qibaipingfei capsule by SBE is better than WE method.

Three strains having degrading poly(?-hydroxybutyrate)(PHB) activity were isolated from activated sludge of different ecological environments and areas,named DS9701, DS9710 and DS9713.The properties of PHB depolymerase produced by DS9701, DS9710 and DS9713 were studied. All the PHB depolymerases are extracellular enzyme and are induced enzyme. The time that enzyme activities of the PHB depolymerases reach the maximum is 96 hours after inoculation. The apparent optimal temperature range for crude enzymes extract is 40℃～45℃.

OBJECTIVE:To study the anti-inflammatory effect of Xinfeng capsule on adjuvant arthritis (AA) in rats. METH-ODS:70 SD rats were randomized into a normal control group(isovolumetric normal saline),a model control group(isovolumet-ric normal saline),a positive control TCM group [Qufeng zhitong capsule 0.4 g(crude drug)/kg],a positive control chemical medi-cine group(Leflunomide tablet 2.1 mg/kg)and groups of low,middle and high doses of Xinfeng capsule,which were respectively marked as groups A,B,C,D,E,F and G. All groups of rats except for Group A were given freund’s complete adjuvant intracu-taneous injection to establishe AA models,and from the 12th day after the inflammation was induced,were given the correspond-ing drug,ig,for 28 consecutive days. The degree of toe swelling and arthritis index were determined and calculated for all groups of rats before and after the administration. The contents of interleukin 1β(IL-1β) and tumor necrosis factor α(TNF-α) in serum and synovial membrane of all groups of rats were determined 24 h after the last administration,and pathomorphological changes of their ankles were observed by light microscope. RESULTS:Compared to group A,group B demonstrated higher degree of toe swelling,arthritis index and contents of IL-1β and TNF-α in serum and synovial membrane,with statistical significance(P<0.01). After the administration,the degree of toe swelling and arthritis index of groups C,D,E,F and G reduced,with statistical signifi-cance(P<0.05). Compared to group B,groups C,D,E,F and G showed lower degree of toe swelling,arthritis index and con-tents of IL-1β and TNF-α in serum and synovial membrane,with statistical significance(P<0.01 or P<0.05). Synovial membrane hyperplasia and inflammatory cell infiltration were noted in the ankle tissue of the rats in group B,and such symptoms were not so serious in groups C,D,F and G. CONCLUSIONS:Xinfeng capsule has anti-inflammatory effect for AA rats by a mechanism which may be related to accelerating synoviocyte apoptosis and inhibiting synovial membrane hyperplasia.

In order to investigate the prevalence and track genetic and antigenic evolutions of infectious bronchitis virus (IBV) and their prevalence in Guangxi, China since 1985, gene amplification and sequencing and virus neutralization (VN) test on chicken embryo tracheal organ cultures were used in genotyping and serotyping of 28 IBV isolates during 2009-2011 in Guangxi. The results of N gene sequencing and comparison showed that the 28 isolates and reference strains were classified into three groups, and most isolates belonged to group Ill, while the isolates in 1985-2008 belonged to groups IV and II. The data of VN test indicated that the 28 isolates belonged to 6 serotypes; among them, 71. 4% belonged to serotypes 1, 2, and 3, and 11 (39.3%) shared the same serotype with the current vaccine strains. Given the data of our previous study, it is found that prevalent serotypes and their proportions varied in different areas of Guangxi and during different periods. These data lay a good foundation for developing an oil-emulsified inactivated polyvalent vaccine containing local dominant serotypes for the effective prevention and control of infectious bronchitis.
Animals ; Antibodies, Viral ; immunology ; Chick Embryo ; Chickens ; China ; epidemiology ; Coronavirus Infections ; epidemiology ; immunology ; veterinary ; virology ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; epidemiology ; immunology ; virology

Retinal degenerative diseases are kinds of leading cause of blindness characterized by photoreceptor apoptosis and progressive neuronal degeneration. A large body of research has shown the evidence of inflammation reaction in such diseases. Retinal neuroinflammation may be a main factor resulting in apoptosis of photoreceptors and neurodegeneration of retina. In addition, the inflammatory response is not only detected in the retina, but also in aqueous humor, vitreous and even in blood, which forms a persistently very low degree, and chronic inflammatory environment. In this review, we focus on the development of microinflammatory response and its predictor for disease outcomes.

Objective To investigate the immunological pathogenesis of immune reconstitution inflammatory syndrome (IRIS) during highly active antiretroviral therapy( HAART), in this prospective cohort study we analyzed the lymphocyte subsets, lymphocyte activation, changes in regulatory T cells, and levels of Th1 and Th2 cytokines in both IRIS and non-IRIS groups. Methods Two hundred and thirty-eight AIDS patients received HAART and participated prospective research cohort for 24 weeks follow-up. Forty-seven IRIS cases and 191 non-IRIS cases were enrolled in the IRIS group or non-IRIS group respectively. Blood samples were collected in both groups at pre- and post-HAART 12 weeks, 24 weeks. Using flow cytometer to detect the immunophenotypes of lymphocyte subsets (CD4 + CD45RA+ CD62L+, CD8+ CD45RA+ CD62L+naive T cells; CD4+ CD45RO+, CD8+ CD45RO+ memory T cells), activated T lymphocytes (CD4+CD38 +, CD8 + CD38 + cells), and regulatory T cell ( CD4 + CD25 + Foxp3 + ). Blood samples collected at pre-and post-HAART4 weeks, 12 weeks, 24 weeks and used ELISA to detect IL-2, IFN-γ, IL-4, IL-10and IL-7 cytokine serum levels. Results The percentages of CD4 + and CD8 + naive T cells and mlemory T cells exhibited no significant differences at the baseline, 12 weeks, 24 weeks of HAART initiation between both groups, but CD4 + and CD8 + memory T cells were demonstrated a trend towards to increase while compared to baseline during HAART. The percentages of CD4 + and CD8 + activated T cells are significantly higher at the baseline while compared to normal control and demonstrated a downward trend, but between both groups showed no significant difference. The percentages of CD4 + regulatory T cell was lower in IRIS group than non-IRIS group at the baseline, 12 weeks, 24 weeks and the onset of IRIS. Th1 cytokines, IL-2 and IFN-γshowed an upward trend during HAART at the levels of IRIS group had significantly increased at 4 weeks and the onset of IRIS. Th2 cytokines, IL-4 and IL-10 showed a downward trend during HAART,and the levels of IL-10 in IRIS group had significantly decreased at 4 weeks and the onset of IRIS. IL-7 was higher than normal control at the baseline in two groups and showed a downward trend during HAART. The level of IL-7 was higher than non-IRIS group at all follow-up points. Conclusion Memory T cells appear rapid increase in the early stage of HAART and may play a significant role in the inflammatory response of IRIS. CD4 + and CD8 + naive T cells, memory T cells and activated T cells showed no significant difference between IRIS and non-IRIS group within 24 weeks after HAART started. There was a significant reduction in the frequency of regulatory T cells in IRIS group without obvious upward trend during HAART, suggesting that the immune suppression function of regulatory T cells in IRIS was impaired. IL-2 and IFN-γ significantly increased while IL-10 significantly decreased at 4 weeks post-HAART initiation and onset of IRIS in IRISgroup than non-IRIS group, suggested that IRIS was related to cytokines environment disorder. That is, a significant increase in inflammatory cytokines, while the relative lack of non-inflammatory cytokines. The level of IL-7 decreased gradually after HAART started, and it was higher in IRIS group when compared to non-IRIS group in the first 24 weeks after HAART started. Also IL-7 may play a role in the pathogenesis of IRIS.

Objective Midkine ( MK ) and nuclear factor-kappa B ( NF-kB ) play pivotal roles in tumorigenesis, which are considered as promising cancer biomarkers. The efficacy of MK and NF-kB as markers for diagnosis and prediction of synchronous metastasis in papillary thyroid cancer ( PTC ) was the aim of present investigation. Methods Seventy six cases of PTC and seventy cases of multi-nodular goiter ( MNG ) were retrieved. The PTC group was further divided into subgroup 1 (16 cases with synchronous metastasis) and subgroup 2 (60 cases without metastases). A retrospective review of clinical information, radiological examinations,131 I treatments and post-131 I-therapy scans were done. Immunohistochemistry of MK, NF-kB p65, and Ki-67 was performed on paraffin-embedded specimens and results were quantified. Diagnostic values of the parameters were conducted by receiver operating characteristic (ROC) curves. Diagnostic sensitivity, specificity, accuracy, positive predictive value, and negative predictive value were determined. Protein levels of MK and NF-kB p65 were then confirmed by Western blot. Results Immunoreactivities of MK and NF-kB p65, and positive percentage of Ki-67 were significantly higher in PTC group than in MNG group (all P<0. 01). ROC showed good differential diagnostic capabilities of all three parameters with diagnostic accuracies of 82. 192% , 80. 137% , and 84. 091%respectively. Moreover, all three parameters were significantly higher in subgroup 1 than those in subgroup 2 (all P<0. 01). ROC showed good predicting efficacies in synchronous metastasis of all three parameters with diagnostic accuracies of 82. 895% , 80. 263% , and 76. 316% respectively. By one-way analysis of variance, Western blot showed that MK and NF-kB p65 protein levels in lesions from subgroup 1 were significantly higher than those from subgroup 2, both were significantly higher than those in MNG lesions ( P<0. 01). Conclusion MK and NF-kB immunohistochemistry can potentially be used for differential diagnosis between PTC and MNG, and for prediction of synchronous metastases.

BACKGROUNDTo study the methods for monitoring nephrotoxicity of cisplatin in lung cancer chemotherapy.

METHODSThe serum urea nitrogen (BUN), creatine (Cr) and urine β2-microglobulin ( β2-MG),α1-microglobulin(α1-MG), albumin (Alb), transferrin (TRF), and retinol-binding-protein (RBP) were measured dynamically in 61 patients with lung cancer who received high-dose chemotherapy with DDP (80- 120 mg/m2) (totally 114 cycles), and some indexes of early nephrotxcity were screened.

RESULTSAt the early stage of chemotherapy (d1-5), the abnormal rates of urine β2-MG and α1-MG were much more predominant than those of other indexes (P < 0.001). The abnormal rates of serum BUN and Cr were increased significantly in the late period of chemotherapy (d10). There were abnormal elevation of urine β2-MG and/or α1-MG at the early stage of chemotherapy for the patients whose serum BUN and creatine were all abnormal at the late stage.

CONCLUSIONSThe urine β2-MG and /or α1-MG might be the valuable indexes for early diagnosis of nephrotoxicity in lung cancer chemotherapy with cisplatin.

Objective To investigate the effect of platelet-rich plasma(PRP) combined with bone graft in the treatment of humeral condylar bone defect.Methods A total of 135 patients with humeral condylar bone defect in Ankang central hospital from January 2012 to December 2015 were divided into the PRP combined group(n =69) and the conventional group(n =66) according to the order of admission time.The patients of PRP combined group were treated with platelet-rich plasma combined with autologous bone graft,and patients of conventional group received autologous bone graft,respectively.The surgery time,hospitalization time,wound healing,fracture union and the motion of elbow joint at postoperative 1 year between two groups were compared.The Kaplan-Meier survival curve was used to reflect the bone healing in both groups,and the log-rank test was used to compare the result.Results There was no statistically significant difference in the surgery time,hospitalization time,wound healing and motion of elbow joint at postoperative 1 year between the two groups(P ＞ 0.05).But the average time of wound healing (3.8 ± 0.72) weeks and the time of bone union (18.8 ± 3.50) weeks in PRP combined group were significantly shorter than (6.4 ±0.58) weeks and (22.7 ± 1.55) weeks in the conventional group(P =0.000),the differences were significant.The KaplanMeier survival curve of the bone union in the PRP combined group was also significantly better than that in the conventional group.Conclusion PRP can promote the healing of fracture in patients with humeral condylar bone defect after autologous bone graft,which contributes to the recovery of elbow function.