Objective To evaluate the efficacy and safety of centella triterpenes cream(R) for treating crow's feet.Methods A double-blind,randomized,vehicle-controlled 12-week study was conducted.Centella triterpenes cream(R) was applied to the lateral canthus on one side (treatment side) and vehicle-(c)ontrol cream to the lateral canthus on the other side (control side) 3 times daily.Efficacy was evaluated based on an investigator-blinded assessment,subject self-blinded assessment and a quantitative analysis by Visioscan(R)VC98 at the baseline,4,8,12 weeks after the beginning of treatment.Results Thirty-six volunteers were recruited and 35 subjects completed the 12-week trial.The investigator-blinded assessment showed a significant difference in the changes of wrinkle scores between the treatment side and control side after 4 weeks (P ＜ 0.05),and the improvement of wrinkles was more obvious on the treatment side than on the control side at 8 and 12 weeks with a statistical difference in the wrinkle scores (both P ＜ 0.05).Compared with the control side,a significant increase in SEw value,which suggested an improvement in wrinkles,was observed on the treatment side after the application of centella triterpenes cream(R).Subjects' assessments revealed no significant difference in the occurrence of irritation or the improvement of coarse wrinkles,whereas the treatment side was superior to the control side in the improvement of skin texture (P ＜ 0.05) at the lateral canthus.Conclusion Centella triterpenes cream(R) thrice daily is effective for the improvement of crow's feet with no obvious side effects.

AIM: To optimize the formulation in the preparation of the Breviscapine Liposomes. METHODS: Using film evaporation-extrusion method to prepare Breviscapine Liposomes according to the uniform design,a optimum formulation was established by determining the entrapment efficiency and the ratio of loading drug. RESULTS: The entrapment efficiency and the ratio of loading drug of Breviscapine Liposomes prepared with cholesterol and lecithin were determined to be 69.60% and 29.06%,respectively.The average diameter is 105.6 nm. CONCLUSION: The application of the uniform design is useful to achieve a large entrapment efficiency and ratio of loading drug.

Objective To observe the growth and biological features of bone mesenchymal stem cells (BMSC) from psoriatic patients. Methods Peripheral blood mononuclear cells were isolated from 5 patients with active psoriasis vulgaris and 5 normal human controls, and BMSC were obtained and purified using plastic adherence method followed by primary culture and passage in vitro. The cell morphology, density and growth were observed with microscopy. Cell growth pattern was evaluated by MTT assay. Flow cytometry was applied to identify surface antigens, including CD29, CD34, CD45 and CD106, on these cells. Results No significant difference was observed in the morphology of primary or descendant BMSC between the patients and controls.The primary BMSC from psoriatic patients tended to adhere to the plastic wall later, confluence and grow more slowly compared with those from the controls. The BMSCs from both psoriatic patients anti healthy donors were positive for CD29, but negative for CD34 or CD45. On the 4th day of culture, the BMSC from psoriatic patients exhibited a decrease in proliferation, with the absorbence at 470 nm (A470) being 0.081±0.0066 and 0.095±0.0130, respectively for BMSC from the patients and controls (t=2.358, P<0.05). Conclusion There is a decrease in the proliferation of BMSC from psoriatic patients which show a morphological similarity to those from healthy controls.

Objective To observe the effects of Buxu Huayu P rescription(BHP)on myeloid hematopoiesis in tumor -b ear-ing mice after combined chemotherap y.Methods Tumor -bearing mouse models were established by implanting P388tu-mor strain and the arrest of myeloid h ematopoiesis in the models was induced by intraperitoneal injection of c yclophos-phamide(CTX)and cytarabine(Ara -c ).The mice were allocated to 3groups:the normal control group,the model group and BHP group.Peripheral bloo d cell count,bone marrow karyocytes(BMK)count,colony count of progenitors o f granulocytes /monocytes(CCP -GM)and the proliferation of bone marrow cells(BMC)were examined.Results The combination of CTX and Ara -c could decrease plasma hemoglobin(Hb )content,the number of white blood cells(WBC),BMK and CCP -GM and the proliferatio n of BMC in model mice(P

Aim To investigate the effects of polyI:C on angiogenesis in mouse prostate carcinoma and its mechanisms.Methods Prostate carcinoma bearing mice were randomly divided into two groups according to tumor volume:contrl group and polyI:C group.After seven times′treatment,the mice were sacrificed.The content of NO in tumor was measured by nitric oxide assay kit.Tumor tissues were partly performed hematoxylin-eosin staining to observe morphological changes and distribution of vasa.Immunohisto chemical staining was used to observe the expression of VEGF,eNOS and AQP1.Results The content of NO in polyI:C group and the control was(1.22?0.77)?mol and(8.73?5.34)?mol respectively,and there was significant difference between two groups(P

Objective To establish an in vitro model of mycobacterial granuloma.Methods Mononuclear cells were isolated from peripheral blood of healthy human subjects,and stimulated to differentiate into macrophages,which were then classified into four groups to be cocultured with Mycobacterium marinum,Mycobacterium tuberculosis,Bacillus Calmette-Guérin,and Mycobacterium leprae,respectively,for five days followed by incubation with peripheral blood mononuclear cells (PBMCs) from the corresponding donors to establish an in vitro model of mycobacterial granuloma.The macrophages cocultured with PBMCs or mycobacteria alone served as the control.Microscopy was performed to dynamically visualize the formation of granuloma in vitro,flow cytometry to detect the expressions of cell surface antigens at different stages,real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA) to determine the mRNA expressions of important cytokines and their protein levels in the supernatant of macrophages,respectively.Results After 7-9 days of coculture with mycobacteria and PBMCs,the macrophages aggregated to form granuloma-like clumps,and some cells fused to form multinuclear giant cells,along with the expressions of some surface antigens such as CD14,CD68 and CD86 on these macrophages.The mRNA expressions of some important cytokines,including tumor necrosis factor-a,interferon-γ interleukin (IL)-1 β and IL-10,were detectable in the macrophages cocultured with mycobacteria and PBMCs,and the secretion of these cytokines was confirmed by ELISA in the supernatant of these cells.Conclusions An in vitro model of mycobacterial granuloma is basically established,which may facilitate the investigation into the formation of granuloma caused by and immune response to mycobacterial infection.

Objective To discuss current detection of food intolerance in health check-up populations and the influence of food intolerance on immune system and inflammation parameters.Methods Serum levels of 14 kinds of food-specific IgG antibodies were analyzed by using enzyme linked immunosorbent assay (ELISA) among 257 healthy check-up adults.White blood cell count,eosinophils,globulin,erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),rheumatoid factor (RF),antinuclear antibodies (ANA) and T cell function were compared between the positive group and the negative group.Student's t test and Chi-square test were used for data analysis.Results Food-specific IgG antibodies were found in 134 participants (52.14％).The most common intolerant foods were egg,crab and milk.Moderate to severe food intolerance was caused by egg or milk.There was significant difference in eosinophils and serum globulin between the positive and the negative group (t =-0.07,x2 =8.91,both P ＜0.05).However,no difference was observed in white blood cell count,ESR,CRP,RF,ANA and T cell function between two groups (all P ＞ 0.05).Conclusion Eosinophils and serum globulin may significantly increased in people with food intolerance,although white blood cell count,ESR,CRP,RF,ANA and T cell function could be normal.

Objective To investigate the effect of erythropoietin (EPO) on mesenchymal stem cells (mMSCs) proliferation under acute kidney injury (AKI) microenvironment,and to study its possible mechanism.Methods C57BL/6 mice's MSCs (mMSCs) were isolated by Percoll density gradient centrifugation and adherence cultivation.Surface markers were identified by flow cytometry.AKI mice models were made by clamping bilateral renal pedicles for 30 minutes and reopening for 30 minutes.Then both renal cortex was drew immediately to make IR kidney homogenate supernatant.P3-mMSCs were divided into different groups: Group A: low glucose DMEM medium with 10% fetal bovine serum; Group B: low glucose DMEM medium with 10% fetal bovine serum plus IR kidney homogenate supernatant; Group C: low glucose DMEM medium with 10% fetal bovine serum plus IR kidney homogenate supernatant and different concentrations of EPO (1,5,10,50 U/ml).Each group was incubated for 1 d,3 d,5 d,7 d.Proliferation of mMSCs was detected by CCK-8,and apoptosis was detected by TUNEL.The protein expression of erythropoietin receptor(EPOR) and the proteins of proliferation/apoptosis related signal pathway were examined by Western blotting.Results Under IR kidney homogenate supernatant,the proliferation ability of mMSCs decreased significantly (P＜0.01),while the apoptoic percentage was significantly higher than that of Group A (P＜0.01).After intervention of EPO,mMSCs proliferation enhanced,at the same time,the apoptoic percentage decreased,in a dose-dependent manner.EPOR was positive in P3-mMSCs by Western blotting.EPO decreased the expression of caspase-3 in mMSCs under AKI microenvironment in a dose- and time-dependent manner,but increased the expression of Bcl-2.Cultured for 5 d,the expression of phosphor-Janus kinase2(p-JAK2) [(0.641 ±0.028) vs (0.456±0.012)] and phosphor-signal transducer and activator of transcription(p-STAT5)[(0.398±0.016) vs (0.209±0.020)] was significantly higher in 10 U/ml EPO group compared to group B.Conclusion Erythropoietin can promote proliferation of mMSCs in vitro under AKI microenvironment,which is mediated by EPOR and related with proliferation/apoptosis signal pathway.

Objective To detect gene mutations associated with dapsone-,rifampicin-and ofloxacinresistance in lesions of patients with recurring or treatment-resistant leprosy collected from 2010 to 2011.Methods Clinical data and lesional specimens were collected during 2010-2011 from patients with recurring or treatment-resistant leprosy who were diagnosed and reported by provincial centers for leprosy control.Mycobacterium leprae DNA was extracted from the specimens and subjected to PCR for the amplification of folP1,rpoB and gyrA genes.The PCR products were directly sequenced and BLAST program was used to compare the sequence of isolated strains with the reference sequence in GenBank.Results Twenty-four patients were enrolled in this study,including 13 with recurring leprosy and 11 with treatment-resistant leprosy.Twenty-one patients showed positive PCR results in all the three regions.Of these PCR-positive specimens,3 from 1 patient with recurring and 2 patients with resistant leprosy harbored a point mutation,acc (threonine)→gcc (alanine),at codon 53 in the floP1 gene,1 from a patient with recurring leprosy harbored a missense mutation,gat (aspartic acid) → aac (asparagine),at codon 441 in the rpoB gene.Conclusions Mutations are detected in the folP1 and rpoB genes,which are associated with the resistance to dapsone and rifampicin respectively,but not in the ofloxacin resistance-associated gyrA gene,in Mycobacterium leprae isolates from patients with recurring or treatment-resistant leprosy.

Objective To investigate the migration of bone-marrow mesenchymal stem cells (BMSCs) under acute kidney injury (AKI) microenvironment in vitro and the effect of erythropoietin (EPO) intervention,and to explore its underlying mechanism.Methods Renal tubular epithelial cells (RTECs) were cultured in hypoxia/re-oxygenation (HR) condition for 12 h,respectively,in order to establish HR-RTEC.BMSCs and RTECs were co-cultured by Transwell system and were divided into 7 groups:control group (group①,only BMSC cultured),BMSC-RTEC co-culturing group (group ②),BMSC-HR-RTEC co-culturing + EPO intervention groups (group ③to group ⑦,EPO concentration:0,1,5,10,50 IU/ml).All the groups were cultured for 48 h and the number of migrating BMSCs was detected.Western blotting was applied for the detection of SDF-1 expression in RTECs and pMAPK and MAPK levels in BMSCs.SDF-1 concentration in the RTECs culture supernatant was tested by ELISA.Results The number of BMSCs migrating to the low chamber where HR-RTECs were cultured was increased,and EPO intervention further enhanced this migration which reached the peak at the concentration of 10 IU/ml [Compared with group③,(46.67±7.37) cells vs (19.00±2.37) cells,P ＜ 0.05].Intracellular expression level and the secreated level of SDF-1 in HR-RTECs in group③ were higher than those in RTECs of group② [0.37±0.01 vs 0.19±0.01,P ＜ 0.05; (61.64±4.88) μg/L vs (35.26±8.78) μg/L,P ＜ 0.05].EPO intervention increased above SDF-1 levels and reached the peak at the concentration of 10 IU/ml [group⑥ vs group③:(173.53± 14.66) μg/L vs (61.64±4.88) μg/L,P＜0.05],accompanied with enhanced phosphorylation of MAPK in BMSCs.Conclusions AKI microenvironment has obvious chemotaxis effect on BMSCs,and EPO intervention can strengthen this effect.The increased SDF-1 level and enhanced phosphorylation of MAPK,the downstream signal protein of SDF-1/CXCR4 axis,are the possible mechanism for EPO performance.