AIM: To study the dynamic levels of tumor necrosis factor alpha (TNF-?), interleukin-6 (IL-6) and gamma interferon (IFN-?) produced by human dendritic cells infected with Dengue virus. METHODS: Monocytes isolated from healthy human peripheral blood were incubated in medium with GM-CSF and IL-4 for more than 7 days. DCs were then collected and identified by scanning electron microscope, immunohistochemistry and lymphocytes stimulatory ability. Human dendritic cells (DC) were infected with Dengue-2 virus (DV-2) in vitro, culture supernatants were collected in different time postinfection (6 h, 12 h, 24 h, 48 h and 72 h), DV antigen in human dendritic cells were demonstrated by an indirect immunofluorescent assay (IFA), production of TNF-?, IL-6 and IFN-? in the culture supernatants were evaluated by ELISA. RESULTS: After 7 days, typical dendritic cells could be obtained. Virus antigen were detected in infected DC by IFA. Dengue virus induces TNF-? and IL-6 secretion from DC and does not induce IFN secretion. CONCLUSION: Human dendritic cells are target of dengue virus infection. TNF-?, IL-6 production from DC are increased with DV infection. Dendritic cells may play an important role in DV pathogenicity and immunity.

Hereditary diffuse gastric cancer (HDGC) is an autosomal dominant inherited disease,and may be related to the mutation of CDH1 or CTNNA1 genes.Microscopically,signet-ring cell carcinoma is suggested frequently in endoscopic biopsy or gastrectomy specimens.Some patients may have concomitant extra-stomach tumor (frequently breast cancer in females).Detection of CDH1 gene mutation should be performed in high-risk individuals,and diagnosis and treatment should be carried out by a multidisciplinary team.Prophylactic gastrectomy is recommended for those with pathogenic CDH1 mutation.Endoscopic surveillance is an option for those with CDH1 mutation of undetermined significance and those without germline CDH1 mutation.This review discussed the concept,genetic characteristics,clinicopathological features and genetic screening of HDGC for providing a reference for clinicians.

Objective To investigate the character and differentiation diagnosis of coronary artery fistula in aged.Methods Clinical symptom, sign and laboratory examination were analyzed in 8 aged patients with coronary artery fistula. Results Myocardial ischemia is the main manifestation of agedness coronary artery fistula, and should be discriminated with coronary artery disease. Conclusion If myocardial ischemia symptom aggravates with age in patient with long-term angina, coronary angiogram is the most dependable method.

The embryo development technique in Zebrafish, Brachydanio rerio, is a toxicity testing method making use of the high sensitivity of fish embryo development in early stage to study and evaluate the specific effecting mechanism, the most sensitive effecting time, embryo toxicity and teratogenicity of chemicals through observing the development process of zebrafish embryo after chemical exposure to fertilized ova. This technique has been widely used to test toxicity of chemicals with the advantages of low cost, high sensitivity, simple to operate and simultaneous to detect multi-endpoints. The main methodology, technical characteristics and the status of world-wide application of this technique are reviewed in this paper. Based on the urgent environmental problems in China, the prospects to use this method for monitoring toxicity of mixed pollutants in wastewater are put forward.

[Objective]To explore the effects of extracorporeal shock waves(ESW) therapy on tendon-bone healing after reconstruction of the anterior cruciate ligament(ACL).[Method]Animal model of reconstruction of the anterior cruciate ligament in 18 healthy F/M New Zealand white rabbits(2～3 months old) were established using digital long extensor tendon and randomly classified into 2 groups.The ESW group was given extracorporeal shock waves therapy on 24 hours after reconstruction of the anterior cruciate ligament while the control group with only reconstruction of the anterior cruciate ligament but no more therapy.All animals were sacrificed at 2、4、8 weeks after operation for histological examinations.The specimens were stained with hematoxylin-eosin and Masson trichrome to investigate pathology change of the tendon-bone healing tissue.The ink intravascular injection was used to observe the neo-vascularization of tendon-bone healing tissue.[Result]The histological examination revealed that at 4 weeks after operation,the ESW group,tendon-bone interface was filled with proliferated fibroblasts and chondroblasts,the collagen fibers were obviously increased and regularly arranged.At 8 weeks after operation,the healing tissue at the bone-tendon interface had developed into dense connective tissue,the collagen fibers were formed in abundance and the perpendicular collagen fibers were regularly and longitudinally arranged.The transition zones of collagen fibers,fibrocartilage and bone(the structure alike a direct ligamentous insertion) occurred in the local region of tendon-bone interface.The histological observations showed that the tissue healing at tendon-bone interface of the ESW group were quicker than those of the control group.The proportion of neo-vessels at the tendon-bone interface of the ESW group were significantly higher than those of the control group at 4 and 8 weeks after operation(respectively P=0.028,P=0.008).[Conclusion]Extracorporeal shock wave treatment can significantly improve the tendon-bone early healing after reconstruction of the anterior cruciate ligament in rabbits.

Objective To study the effects of podophyllotoxin solid lipid nanoparticles (POD-SLN) on the proliferation of human epidermal cells in vitro. Methods POD-SLN was prepared by using microemulsion technique, the morphology of POD-SLN was examined by transmission electron microscope (TEM), and its particle size and Zeta potential were studied by Zetasizer analyzer. High-performance liquid chromatography (HPLC) was employed to determine the entrapment efficiency of podophyllotoxin (POD) in the nanoparticles, and its stability was observed. Human epidermal cells were treated with different concentrations (0.1-1 000 ?g/L) of POD-SLN, and the proliferation of human epidermal cells was studied at different time points after exposure (6h, 12h, 24h, 48h). The cytotoxic effects of POD-SLN, POD liposome, free POD, blank solid lipid nanoparticles (SLN) and control groups on human epidermal cells were assessed using a colorimetric MTT cell viability assay. Results POD-SLN displayed spherical or elliptical in shape, and it was stable. The average particle size of POD-SLN was 87.2?10.3nm, Zeta potential was 25.3?0.8mv and the entrapment efficiency of POD in the nanoparticles was 83.2%?2.5%. POD-SLN inhibited the proliferation of human epidermal cells in a concentration- and time-dependent manner. At the same concentration, the effect of PDP-SLN on anti-proliferation was stronger than that of POD liposome and POD. The inhibition of human epidermal cells after 48h exposure to PDP-SLN, POD liposome, and POD reached 91.05%, 77.02% and 68.46% respectively, at the highest concentration of 1000?g/L, and the IC50 were 2.11?g/L, 16.65?g/L and 101.42?g/L, respectively. Blank SLN had no effect on the proliferation of human epidermal cells. Conclusion This formulation and technology are stable and practical. POD-SLN can significantly inhibit the proliferation of human epidermal cells in vitro and the inhibitory effect was better than that of POD liposome and POD.

The changes of cytochrome oxidase activity of retinal ganglion cell (RGC) caused by the artificial high intraocular pressure (HIOP) was presented in this paper. Normal saline was injected into the anterior chamber of the right eye of 36 rats. that resulted in an intraocular pressure up to 60 mmHg for 3 hours. The experimental rats were divided into 0-, 3- and 7-day groups according to their survival time. Both retinae of each rat were whole mounted on the same slide, the left one being used as control. Thirty-six pairs of retina were treated with the modified cytochrome oxidase (CCO) histochemical technique under similar conditions. The high CCO activity of RGC were counted. The extinction of the cytoplasm of RGC, which indicates the degree of CCO activity, was measured with microphotometer. As compared with the normal eyes, the high CCO activity RGC of experimental eyes were reduced significantly. It was found that the high CCO activity of RGC in the temporal side of retina has been reduced much more than that of the nasal side. However, the high CCO activity of RGC in 3- and 7-day groups were more than that of 0-day group, the extinction of CCO activity of RGC in 7-day group was lower than that of the 3-day group. These facts showed that the CCO activity might restore in various degrees followed by a longer survival time. This experiment emphasized the fact that the HIOP led to metabolic changes of RGC, which may be of value to the study of glaucoma.

Objective To elucidate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) in patients with chronic hepatitis B (CHB), and to explore the correlation between hepatitis B virus (HBV) persistent infection and TLR3 expression. Methods Sixty CHB patients (CHB group) and 20 healthy controls (control group) were enrolled. The peripheral blood mononuclear cells (PBMCs) were isolated and CD14~+ monocytes were sorted by immunomagnetic beads. Immature DCs (imDC) were induced and proliferated in vitro and mature DCs (mDC) were obtained after the poly I:C stimulation. The expression of intracellular TLR3 mRNA was detected by real-time polymerase chain reaction (PCR), and surface markers [CD80 and human leucocyte antigen (HLA)-DR] were determined by flow cytometry after 48 h of stimulation. The comparison of quantitative data was done using t test. The qualitative data were compared using chi-square test.Results The mean fluorescence intensities (MFI) of intracellular TLR3 of imDC before poly I:C stimulation in CHB group and control group were 1212.05 ± 250.80 and 1192.95 ± 301.40,respectively, which were not significantly different (t = 0. 280, P>0. 05). While after stimulation,those were 1352.98± 313.67 and 1593. 00± 349. 65, respectively, the latter was significantly higher than the former (t = 2. 880, P<0. 05). The levels of TLR3 mRNA inside mDCs in both groups were increased after poly I:C stimulation, which were 0. 1204 ±0.0267 and 0. 1780 ± 0.0664, respectively in CHB group and control group, and that in control group was significantly higher (t = 3. 909, P<0.05). Furtherly, patients in CHB group were divided into HBeAg(+ ) and HBeAg( -) subgroups.After stimulation, the MFI and mRNA of TLR3 inside mDC were greatly elevated in both subgroups,but there were no difference between these two subgroups (t = 0. 366, P>0. 05). Conclusions The intracellular expressions of TLR3 in mDC in CHB group and control group are obviously increased after the poly I:C stimulation, but the increased level in CHB group is lower than that in control group. The results suggest that the insufficiency of TLR3 synthesis may be related to the HBVpersistent infection.

Objective To detect the expression of type Ⅰ interferon in monocyte-derived dendritic cells(MoDCs)after Toll like receptor(TLR)3 triggered in patients with chronic hepatitis B(CHB),and to evaluate immune responses of CHB patients and its roles in the mechanisms of persistent infection of hepatitis B virus(HBV)and chronicity of hepatitis.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and purified using magnetic beads(plasma was saved simultaneously)from 26 CHB patients and 18 healthy volunteers(HV).Dendritic cells(DCs)were induced and proliferated in a culture medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin(rhIL-4).EX3s were stimulated with Poly Ⅰ:C and the supernatants were collected at 0 h and 24 h after stimulation.Type Ⅰ interferon(IFN-α and IFN-β)in plasma and supernatants were examined by enzyme linked immunosorbent assay (ELISA).Results The levels of type Ⅰ interferon in plasma were not significantly different in groups of HV and CH B.IFN-α and IFN-β expressions in supernatants before Poly Ⅰ:C stimulation were(80.00±16.15)ng/L,(36.39±13.90)ng/L in CHB group and(76.76±15.90)ng/L,(37.14±13.68)ng/L in HV group,respectively.And there were no statistical differences between two groups(t=1.651,t=0.178;both P＞0.05).IFN-α expressions in supernatants at 24 h after stimulation in two groups were both higher than those before stimulation(at 0 h),but there were no statistical differences(t=1.534,t=1.243;both P＞0.05).IFN-β expressions in supernatants at 24 h after stimulation in HV group was(54.57±16.80)ng/L,which was significantly higher than that at 0 h(37.14±13.68)ng/L(t=4.061,P＜0.05).However,there was no significant difference at 24 h than tht at 0 h in CHB group(t=1.796,P＞0.05).At 24 h after stimulation.IFN-β level was(54.57±16.80)ng/L in HV group,which was significantly higher than that[(41.64±12.57)ng/L]in CHB group(t=2.921,P＜0.05).Conclusions Functions of MoDCs from CHB patients are impaired and MoDCs could not express type Ⅰ interferon normally.Expression of type Ⅰ interferon after TLR3 triggered in CHB patients is mainly IFN-β.

Objective To study and evaluate the clinical effect of laparoscopic therapy in endometriosis com-plicated with polycystic ovary. Methods To retrospectively analyze 33 cases of endometriosis complicated with poly-cystic ovary and observe the results of symptomatic relief rate and pregnancy rate after laparoscopic therapy. Results The postoperative eumenorrhca rate, dysmenorrhea relief rate, auto-ovulation rate and pregnancy rate were 90.9%, 100% ,87.9% and 60.6% respectively. Conclusions The clinical symptoms of endometriosis complicated with polyeystic ovary are not predominance. This study confirms the laparoscopic therapy in endometriosis complicated with polycystic ovary is an effective approach to increase the symptomatic relief rate and pregnancy rate.